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Olmix SymposiumSeptember, 10th 2012
FRANCE - Palais des Congrès of Pontivy
How Algae can bring solutions to GlobalNutrition & Health issues
«Algae: The Blue Revolution
for a Sustainable Chemistry»
This event is organized by Olmix with the support of its partners:
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Dear partner,
OLMIX, pioneer in algae use for animal nutrition since 1995 has the pleasure to invite you to attend the 1st olmix algae symposium dedicated to “global nutrition and health issues”.
Using algae extract for their polysaccharides and trace-elements materials became com-mon for Olmix in the beginning of the second millennium.
It all started when OLMIX‘s R&D team had the feeling that they could increase the binding potential of Clay by including inside algae polysaccharides. The idea was to increase the interlayer space of natural montmorillonite clay thanks to the ulvans, green algae poly-saccharides. A full program was then developed with CNRS (National Center of Scientific Research) and CEVA (Center for study and valorization of algae) and gave birth to Ama-deite®, the algae based revolutionary and worldwide patented hybrid material. The first great commercial success using Algae in animal feeding was born: MTX+.
These successes encouraged OLMIX to increase its use of Algae for animal and vege-tal health and nutrition. Today ULVANS, a new R&D program, is aimed to provide even more Algae based solutions to the field with more technology inside, using enzymatic hydrolysis and separation techniques from harvest to the final product.
How Can Olmix innovate so much in Algae use? Its location, based right in the middle of a region of the world where most of the Algae scientific knowledge, supply and diversity is concentrated: BRITTANY.
To share this Algae Blue revolution for a sustainable Chemistry with its partners OLMIX organize on September 10th the 1st OLMIX ALGAE SYMPOSIUM “How algae can bring solutions to global nutrition and health issues”. Speakers from the
most recognized specialized institutions will share with us the latest knowledge on Algae science and yet more…
On behalf of Hervé Balusson, OLMIX CEO, we are looking forward to welcoming you soon in Brittany.
Olmix team
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Morning sessions under the Chairmanship of Catherine Boyen Director of the “Marine plants and biomolecules” laboratory - Roscoff Biological Station- CNRS UMR 7139 - Steering committee of Biogenouest®
8.30 Welcome reception
9.00 Welcome speechHervé Balusson Olmix Group President and CEO
Session 1 Algae: A new world to discover
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Introduction - Brittany, historic region in algae valorisation• History of algae situation in the region• Sea World professional organizations and institutes - Current actionsDr. Christine Bodeau-Bellion Science et Mer Laboratory - President of the Syndicate of Seaweed and Marine Plants - Le Relecq Kerhuon
9.30Brittany an area of excellence in algae knowledge• Research organizations in Brittany - Their expertise• Present and future research programsPr. Eric Deslandes University of Western Brittany - Brest
9.50Discovering the Blue Chemistry• General information on algae, their origin and biological characteristicsDr. Philippe Potin Research Director - Roscoff Biological Station - CNRS UMR 7139 - IDEALG project coordinator
Algae: A sugared treasureDr. Mirjaml Czjzek Research Director - Roscoff Biological Station - CNRS UMR 1931
11.00 Coffee break
Session 2 Algae in the service of Health
11.30Enzymatic hydrolysis in chemistry of seaweedsPr. Nathalie Bourgougnon LBCM (Biotechnology and Marine Chemistry Laboratory) - University of Southern Brittany - Vannes
PROGRAM
«Algae: The Blue Revolution for a Sustainable Chemistry»
10.20
How Algae can bring solutions to Global Nutrition & Health issues
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12.00Bioactivities of Marine Polysaccharides in human and animal health (Update)Dr. Henri Salmon Research Director - INRA Tours - Nouzilly (French National Institute for Agricultural Research)
12.30Marine lipids in amplifying cancers chemotherapyPr. Philippe Bougnoux Oncologist, Director of the Unit «Nutrition, growth and cancer» - INSERM(National Institute for Health and Medical Research) - Chief of cancerology service - CHU Tours(University Hospital Center)
13.00 Lunch (marine buffet)
Afternoon sessions under the Chairmanship of Dr. Christine Bodeau-BellionScience et Mer Laboratory - President of the French Syndicate of Seaweed and Marine Plants - Le Relecq Kerhuon
Session 3 Industrial applications originating from algae chemistry
14.15The algae industry in ChileEliana Henriquez Flores Agronomist - Head of the International Affairs Unit - CIREN(Centre for Renewable Natural Resources Information) - Santiago, CHILE
14.30 Algae, source of active principles in cosmeticsAlexis Rannou Deputy Managing Director in charge of Innovation - ARD Soliance - Pommacle
14.50Algae, source of nutriments for humansDr. Maria Hayes Scientific Project Manager - NutraMara - Teagasc Ashtown Food ResearchCentre - IRELANDChristine Le Tennier Algues de Bretagne - Globe Export SARL - Rosporden
15.25 Algae in the service of soils nutritionDr. Bruno Daridon Research and Development Director - PRP Technologies - Paris
16.00 Coffee break
Session 4 Industrial applications originating from algae chemistry (follow)
16.15 Algae in the service of terrestrial plants healthDr. Adeline Picot Plants Pathology Laboratory - VEGENOV BBV - St Pol de Léon
Algae, source of nutrients in animal nutritionPr. Simon Davies Professor of Aquaculture Nutrition at the University of Plymouth (UK).Member of World Aquaculture Society
17.10 Round table: From research to industrial application.
18.30 Cocktail: Marine algae in the castle20.00 Gala dinner and evening festivities - Pontivy castle - Palais des Congrès
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GUEST SPEAKERSIntroduction and presentation
Mr Hervé BalussonOlmix Group President and CEO
Hervé Balusson is the founder and Chairman of Olmix Group, specialized in the trace-elements feed additives and organic fertilizer. He has succee-ded in bringing Olmix from a regional stage into international coverage.Olmix products are now available in more than 50 countries.
Ms Catherine BoyenDirector of the “Marine plants and biomolecules” laboratory - Roscoff Biological Station- CNRS UMR 7139 - Steering committee of Biogenouest®.
Dr. Christine Bodeau-Bellion Science et Mer Laboratory - President of the Syndicate of Seaweed and Marine Plants - Le Relecq Kerhuon
Pr. Eric DeslandesUniversity of Western Brittany - Brest
Dr. Philippe PotinResearch Director - Roscoff Biological Station - CNRS UMR 7139 - IDEALG pro-ject coordinator
Philippe Potin (49), Docteur en biologie, HDR, Directeur de Recherche 2ème classe au CNRS depuis oct. 2006 (SBR, UMR 7139 CNRS-UPMC-Paris6)Dr. Philippe Potin, marine biologist and biochemist has obtained his Ph.D. from the University of Brest in 1992 and continued his post-doctoral research at the NRC Institute for Marine Biosciences in Halifax (NS Canada) and was hired by CNRS in Roscoff. P. Potin’s scientific interests (>70 primary publications) are in the bases of pathogen defense reactions and signaling in marine algae, with an emphasis of the specific traits of marine plants such as the halide metabolism. Research in his team investigates fundamental processes underlying interactions between seaweeds and pests. He was also interested in technology transfer with the Goëmar Laboratories, to develop the use of oligosaccharides for disease control in agricultural crops (4 patents, one product on the market) and during his mandate as a project manager for
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GUEST SPEAKERSIntroduction and presentation
the marine biotechnologies at the Maritime Cluster “Pôle Mer Bretagne”. He is currently the scientific coordinator of IDEALG, a 10-year national integrative pro-ject, within the framework of the French Stimuli Program Investis- sements d’Avenir, to capitalize on the recent breakthroughs in algal genomics to develop seaweed genetics and biotechnology.Philippe POTIN, Station Biologique de Roscoff, BP 74 - 29680 Roscoff, Tel.33-2. 98.29.23.75, Fax.33-2. 98.29.23.85, Mail potin@sb-roscoff.fr
Dr. Mirjaml CzjzekResearch Director - Roscoff Biological Station - CNRS UMR 1931
Mirjam Czjzek has studied chemistry at the University of Frankfurt, then at the TH Darmstadt in Germany where she has obtained her PhD in crystallography. After one year of a post-doctoral position in the ‘Labo-ratory for crystallography of biological macromolecules’ (LCMB) of Mar-seille, she has been recruited at the CNRS in October 1992. She is crys-tallographer by education and has started to work on CAZymes solving the crystal structures of cellulases and beta-glucosidases during several years in the group of Bernard Henrissat in Marseille, France. In 2005 she moved to the Station Biologique de Roscoff, where she is now ‘directrice de recherche’ of CNRS in the laboratory for ‘Marine plants and biomolecules’. Her research program entitled ‘Marine Glycobiology’ currently focuses on the structures and functions of carbohydrate-active enzymes, including their CBMs, which are involved in marine algal cell wall polysaccharide depolymerization.
Pr. Nathalie BourgougnonLBCM (Biotechnology and Marine Chemistry Laboratory) - University of Sou-thern Brittany - Vannes
Nathalie Bourgougnon has been working in the Laboratoire deBio-technologie et Chimie Marines (LBCM) at Université de Bretagne-Sud since 2001. Previously, she was lecturer at the University de la
Rochelle during 8 years. The principal thematic of her research relates to the search for marine substances with biological activities mainly extrac-ted from algae. She has a good experience in the field of the extraction, purification, characterization and evaluation of biological (antiviral, antifouling, antiprolifera-tive…) activities of marine compounds. She has published ca. 55 papers in peer-reviewed journals and book chapters, in particular on antifouling or antiviral substances extracted
from seaweeds, extraction and purification of bioactive marine substances. She has deposit two patents about antiviral substances. She has been involved in
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several Euro- pean projects (e.g. FP4 Bioactive Marine Natural Products in the Field of Antitumo- ral, Antiviral and Immunomodulant Activity, MAST III; FP5 AVINSI- Anti Viral Infection Non Specific Immunity: Basis of non specific immunity against viral diseases in aquacultured species; FP6 Valbiomar Biotechnologique valorization of the marine resources; FP7 Biotecmar: integrated transregional project for communication, technical information and technology transfer in the domain of biotechnological exploitation of marine products and by-products) and recently in OSEO program ULVANS. She has coordinated the project ASEM-DUO from MAE between France and Malaysia (2007-2009). She is member of several networks, Two French networks: BioChiMar concerning marine substances with biological activity and SEAPro (Sustanaible Exploitation of Aquatic PRO-ducts) concerning biotechnological up-grading of fish, seaweeds or aquaculture by-pro-ducts; and an international network: RAQ Quebec Aquaculture Network. At the national level, Nathalie Bourgougnon has collaborated with Dr. JL Mouget (Université du Mans) for physiological approaches of antifouling substances extracted from seaweeds, V. Stiger (Université de Bretagne Occidentale) for marine substances extracted from red seaweeds, Dr T. Renault (IFREMER, La Tremblade) for defences mechanisms of oysters against bacteria and viruses , JP Bergé (IFREMER, Nantes) for upgrading of marine resources and at the international level, she gained experience in cooperation with Morocco (University Tétouan; Pr. H. Riadi), Institute of Marine Biotechnology of University Malaysia Terengganu (Pr. Effendy) for biological compounds extracted from seaweeds, ISMER (Québec, Dr. R. Tremblay) for biological compounds extracted from seaweeds, microalgae, invertebrates. She is Vice-president of International PhD School (Coordination of Doctoral program) of the Université européenne de Bretagne (UEB) www.ueb.eu. At University de Bretagne-Sud, she is in charge of research program and Coordinator of master «Biotechnology» (www-lbcm.univ-ubs.fr).
Dr. Henri SalmonResearch Director - INRA Tours - Nouzilly (French National Institute for Agricul-tural Research)
Dr. Henri Salmon is a Research Director in the Institute of National Agro-nomic Research (INRA) in France. He earned his DVM from the National Veterinary School-Alfort in Paris and his PhD in Immunology from the University of Paris.Prior to joining INRA, he served 6 years as a Research Assistant at Col-lege of Veterinary Medicine in Alfort. Since 1984, he has served as Director of Research, INRA, laboratory of Animal Infectiology and Public Health, Tours-Nouzilly. He served one year in Transplantation Research Biology Center, Harvard Medical School and Massasuchetts General Hospital (Boston, MA).
GUEST SPEAKERSIntroduction and presentation
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GUEST SPEAKERSIntroduction and presentation
The objective of his research is an understanding of the interrela- tions («immune links») between digestive, pulmonary and mammary mucosal im- mune responses to improve protection against pathogens. He has dissected the mechanisms under-lying the migration of IgA plasma cells from the mucosae to the mammary gland in the sow ; these mechanisms substantiate the production of IgA in colostrum and milk. and hence are responsible of passive mucosal protection of the suckling piglets. Now he is looking at the means to shorten the onset of IgA response in gut of weaned piglet. To replace the anti-biotics-growth factors in food, he designed «immunoprobiotic», as vectors to enhance the neonatal gut immunity which deliver enhacing factors of IgA immune response including pre- and probiotics.
Pr. Philippe BougnouxOncologist, Director of the Unit «Nutrition, growth and cancer» - INSERM (Natio-nal Institute for Health and Medical Research) - Chief of cancerology service - CHU Tours (University Hospital Center)
Philippe Bougnoux is a medical oncologist, specialized in breast and gynaecologic cancers. He performed his trainings in medicine in Tours and in immunology at the Pasteur Institute in Paris. After a 3 years post-doctoral staying as a Fogarty fellow at the National Cancer Institute in Bethesda, MD, he became professor of cancer biology at the university of Tours, and chief of the cancer outpatient unit at the university cancer centre Henry S. Kaplan. He belongs to the Inserm research Unit 1069 « Nutrition, growth and Cancer » and has been coordina-ting a consortium of research units in chemistry and biology on marine-derived anticancer agents within the canceropôle of the western part of France, which he heads now. His research interests are to understand how diet and lipid nutrients influence the molecu-lar alterations which result in malignant tumors and how they integrate to delay breast can-cer occurrence or individual response to anticancer agents. He does translational research
in the field of dietary lipids in relation to breast cancer prevention and treatment. He is currently carrying out randomized clinical trials of dietary intervention with omega-3 poly-unsaturated fatty acids to enhance the sensitivity of tumors to radiation or chemotherapy.
Address: INSERM U1069, Henry S. Kaplan Cancer Centre, University Hospital Breton-neau, 37044 Tours, FranceTelephone: +33 (0) 2 4747 8261Email: philippe.bougnoux@inserm.frWeb site: www.n2c.univ-tours.fr
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Ms Eliana Henriquez FloresAgronomist - Head of the International Affairs Unit - CIREN (Centre for Re-newable Natural Resources Information) - Santiago, CHILE
Agronomist - Head of the International Affairs Unit - CIREN (Centre for Renewable Natural Resources Information) - Santiago, CHILEIn the period between the years 2004-2008, working as Chief of Labora-tory Sub-department and Quarantine Agricultural Station, from the Agricultural and Lives-tock Service, she has made an important management of inter-agency cooperation at the national and international level with different research institutions of great renown and reco-gnized academic prestige around the world. This has enabled that SAG, at present, has signed various «Memorandum of understanding» or Agreements of International Coope-ration with institutions of Spain, Scotland, Italy and United States. Thus, all the necessary efforts were made that will, in the near future, allow to establish agreements with England, Australia, France and New Zealand.The technological horizon for Laboratory Sub-Department and Agricultural Quarantine Station was expanded through the emphasis on inter-agency, both national and foreign cooperation. In this way, one can access to the techniques implemented and developed in important centers of research around the world
M Alexis RannouDeputy Managing Director in charge of Innovation - ARD Soliance - Pommacle
Ingénieur Agricole (ISAB) 1991Ingénieur d’études ARD (Agro industrie Recherche et développement) en charge de la sélection variétale de la betterave biotechnologie pour la fabrication d’acide galacturonique.1994 Responsable du pilote industriel ARD mise au point de tension-actifs verts (Uronate de sodium et Alkyl polypentosides)1997 Directeur technique SOLIANCE & développement industriel Amadéïte avec OLMIX2000 Formation IFG CGDPME (Gestion des entreprises) 2002 Directeur Général Adjoint en charge de la production et du compte l’Oréal2007 DGA en charge de l’innovation (8 Brevets) 20 bx produits et du marketing stratégique2012 DGA en charge de l’innovation Soliance et Wheat Oléo
GUEST SPEAKERSIntroduction and presentation
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GUEST SPEAKERSIntroduction and presentation
Dr. Maria HayesScientific Project Manager - NutraMara - Teagasc Ashtown Food Re-search Centre - IRELAND
NutraMara Scientific Programme Manager & Principle Investigator - Work Package 2 and 7.Main Research Interests: • Isolation, purification and characterisation of marine derived molecules, especially peptides and phlorotannins from marine seaweeds and by-products • Fermentation • Bioassay development with a particular focus on heart and mental health disorders – i.e., renin, ACE-I, PAF-AH, PEP and inhibition of other enzymes with heart and mental health effects • Generation of chitin and chitosan from marine shellfisheries waste streams • Isolation and characterisation of enzymes (in particular chitinolytic enzymes)Short Biography:Dr Hayes obtained her BSc (Hons) in Science, specialising in Industrial Microbiology and Chemistry from University College Dublin (UCD). She carried out her PhD at the Teagasc Food Research Centre, Moorepark and University College Cork in the area of bioactive peptide isolation and characterisation from milk proteins and waste streams (whey and casein). She then carried out Post-doctoral work at the Centre of Applied Marine Biotech-nology in Donegal where she worked on the isolation of chitinolytic enzymes from shell-fisheries crab and whelk waste streams. She is currently the NutraMara Scientific Pro-gramme Manager and supervises two NutraMara PhD researchers who are funded by the Teagasc Walsh Fellowship programme. These students are Mr Ciaran Fitzgerald and Ms Michelle Tierney.
Selected publications: Fitzgerald, C., Gallagher, E., Tasdemir, D., Hayes, M., (2011), Heart Health peptides from
macroalgae and their potential use in functional foods. Journal of Agriculture and Food Chemistry, DOI: 10.1021/jf201114d Di Bernardini, R., Harnedy, P., Bolton, D., Kerry, J., O’ Neill, E., Mullen, A. M., Hayes, M., (2011), Antioxidant and antimicrobial peptidic hydrolysates from muscle protein sources and by-products. Food Chemistry, 124, 1296-1307.
Tierney, M. S., Croft, A. K., Hayes, M., (2010) A review of antihypertensive and antioxidant activities in macroalgae, Botanica Marina, 53 (2010), 387-408.
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Hayes, M., Carney, B., Slater, J., Bruck, W., (2008), Mining marine shellfish wastes for bioac- tive molecules: Chitin and chitosan; Part B: Applications. Biotech-nology Journal, 3, 7, 871-877.Hayes, M., Barrett, E., O’Connor, P., Gardiner, G., Fitzgerald, G., Hill, C., Stanton C., Ross R.P. (2007), Salivaricin P: one of a family of two component anti-listerial bacteriocins pro-duced by intestinal isolates of Lactobacillus salivarius, Appl Environ Microbiol. 73, 11, 3719-3723.
Ms Christine Le TennierAlgues de Bretagne - Globe Export SARL - Rosporden
«“I was born an entrepreneur”Christine Le Tennier is a dynamic person. An impulsive one. She is com-plete. Political cant, she does not know it. “I was born an entrepreneur.” A witticism? Not at all. Before she was 20 years old, Christine Le Tennier did not have any idea about what the wage system was. “My grandpa-rents were corporate managers.” Farmer on her paternal side, tinsmith in Alger for her maternal grandmother. When she was 20, she was hired by Hilton. As a barmaid. At age 22, she became a commercial executive. Still at Hilton. In Ontario – where she was born – and in New York State. When she was offered a big job in Africa, she turned it down. Went back to Brittany, met her future husband and went back to school to study international business.In 1986 Christine Le Tennier created Snc Glob’export with the aim of international consul-ting and trade. Globe export became Sarl Globe Export – Seaweed of Brittany in 1993, opening date of the first production factory of seaweed-based products.Today edible seaweed are lacking in Brittany, studies foresee a field for seaweed in 10 years. “But I do need seaweed here and now.” Meanwhile the development of this field, Christine Le Tennier imports a part of seaweed she transforms and makes a turnover of 2 million Euros with 13 to 15 employees. Customers of Seaweed of Brittany: industry, catering, retail, export (20%), mail order selling. The strategy developed in 2012 can be summarized in 2 major divisions: innovation and interna-tional.
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GUEST SPEAKERSIntroduction and presentation
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GUEST SPEAKERSIntroduction and presentation
Dr. Bruno DaridonResearch and Development Director - PRP Technologies - Paris
Bruno Daridon, 51 years old, integrated PRP Technologies in March 2007 as R&D manager, and then joined the executive committee. He is an agronomist (ENSAIA Nancy-1984) and Doctor in Biotechnology and Food Process Engineering (INPL Nancy- 1988). From 1993 to 1997, he created as a R&D engineer, Prabil S.A., a society of research on hire about extraction and functionalization of plant molecules and valuations of non-food agricultural products, then became its general manager in 1997. From 2004 to 2007, he was in charge of the site Novasep Brabois where he developed processes of fractionation and purification of biomolecules for the pharmaceutical industry.
Dr. Adeline PicotPlants Pathology Laboratory - VEGENOV BBV - St Pol de Léon
After graduating with a PhD in plant pathology from the University of Paris-Sud 11 in 2010, Adeline Picot has been working at Vegenov as a Plant pathology assistant for one year. Her field of research focuses on the evaluation of plant defense elicitors and the optimization of their use in several pathosystems including grey molds and powdery and downy mildews in tomato, strawberry… She is involved in the French network Elicitra which aims at understanding, developing and promoting the strategy of plant de-fense elicitors.
Pr. Simon DaviesProfessor of Aquaculture Nutrition at the University of Plymouth (UK). Member of World Aquacul-
ture Society.
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La Bretagne : Terre historique de la valorisation des algues
Christine BODEAU
Présentation dédiée à : • Jean DUGOUJON, créateur de la Chambre Syndicale des Algues et Végétaux Marins • Pierre ARZEL, Chercheur à IFREMER • Jean-Yves FLOCH, Professeur à l’Université de Bretagne Occidentale
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3 entreprises dans années 1960’
21 entreprises aujourd’hui : diversité des applications
La Chambre Syndicale des Algues et Végétaux Marins
Bret’algue
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Source : Google Earth
Source : Google Earth
La Bretagne : Une situation géographique unique
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Source : Google
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Jusqu’à 13 m de marnage dans la Manche !
Marnages dans le Monde. - Source : SHOM Plus petits = verts / Plus grands = rouges
Les algues se répartissent sur tout l’estran :
La Bretagne : Une région au marnage exceptionnel
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Les débuts de la récolte en bateau
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La récolte à pied
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M.Philippe
La récolte à pied
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L’algoculture
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Le séchage
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Le brulage
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Extraction d’iode
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Les utilisations industrielles et domestiques
XVIIème fabrication du verre
Engrais
Début XIXème, fabrication de l’iode (jusqu’en 1955)
Nourriture du bétail (depuis le XIXème)
Literie
Épaississant et gélifiant : alginates début du XXème / carraghénanes 1960
Alimentaires, légumes de la mer, 1980 (essor)
Cosmétique, pharmaceutique
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Molécules à rôle physiologique
de survie : antibiotiques,
antioxydants…
Oligoéléments Sels minéraux
Polysaccharides Sucres
osmolytes
Algues et biochimie
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Oligoéléments Sels minéraux
• Engrais • Cosmétiques • Thalassothérapie • Alimentation • Diététique (calcium) • Médecine (iode) • Nutrition animale
Algues et biochimie
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Polysaccharides Sucres
Alginates (Algues brunes)
Carraghénanes (Algues rouges)
Laminarine (accélérateur de
croissance)
Produits épaississants et
gélifiants
Engrais
Divers Pharmacie
Alimentation Diététique
Algues et biochimie
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium Algues et Mer ©
Molécules à rôle
physiologique de survie :
antibiotiques, antioxydants…
Molécules à haute valeur
ajoutée
Algues et biochimie
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Bord à Bord ©
Algues de Bretagne©
Alimentaire
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La récolte aujourd’hui en bateau
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Discovering the Blue Chemistry of algae:
Their origin, biology and metabolism
Philippe POTIN - Team Algal Defenses CNRS-UPMC UMR 7139 - Marine Plants & Biomolecules
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Seaweeds from
Roscoff: an exceptional
place for
studying biodiversity
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Seaweed belts on the shore
3 Floc'h, J.-Y. (1964). Distribution verticale et écologie des algues marines sur les côtes Bretonnes. Penn ar Bed 4(37)
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Reds and greens
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Phaeocystis globosa
Brown’s
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Modern stromatolites in Shark Bay, Western Australia http://en.wikipedia.org/wiki/Stromatolite
2.724 billion years ago as far back as 3.450 billion years ago
Blue green algae
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Algae are photosynthetic organisms. Based on the pigment and food reserve, algae are classified into different types, namely, blue green algae (BGA), green algae, red algae and brown algae.
Algae origins are not only a matter of pigments
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The tree of life (Haeckel, 1866, c/o Simonetta Gribaldo).
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium Lane et al, TREE, 2008
Algae in the eukaryotic tree
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Endosymbiotic gene transfer: organelle genomes forge eukaryotic chromosomes Timmis et al. (2004) Nature Reviews Genetics 5, 123-135
The endosymbiotic origins of eukaryotes
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between 2 and 1.5 billion years
Endosymbiotic gene transfer: organelle genomes forge eukaryotic chromosomes Timmis et al. (2004) Nature Reviews Genetics 5, 123-135
The endosymbiotic origins of eukaryotes
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? about 1,2 billion years
Secondary endosymbiotic origin of brown’s
12 The life of diatoms in the world's oceans E. Virginia Armbrust Nature 459, 185-192
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Secondary endosymbiotic origin of other algae
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Seaweeds belong to independant lineages
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium Dr Kathleen Drew-Baker
Porphyra life cycles (1949)
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Porphyra aquaculture in Asia (nori)
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Ulva life cycles
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Kelp forests and life cycles
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Kelp aquaculture
Source: Ifremer 19
Olmix Algae Symposium - Sept. 10th, 2012
www.seaweedenergysolutions.com/ -
Saccharina latissima farming
Kelp aquaculture
20 Olmix Algae Symposium
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Algal metabolisms
21
25
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Kelp biorefinery
22 Find an alternative to the storage of fresh biomass in formaldehyde
Acid Calcium
Na2CO3
LAMINARIN MANNITOL
FUCANS Na- ALGINATE
Co-extraction of laminarin, mannitol , fucans with alginates
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Iodine first discovered in 1811 by Courtois in kelp ashes
Production of « soda » bricks by seaweed harvesters by burning dried kelps in stone ovens
Kelps : a major source of iodine
23
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Iodine may serve as an inorganic antioxydant in kelps Küpper,. et al. (2008) Proc. Natl. Acad. Sci. USA 105, 6954-8
24
26
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
P. Rouzé / Y. van der Peer
+ Es Genome Consortium
Brown algal genomics
GENOME SEQUENCING: 3,000,000 reads (10X, shotgun)
cDNA SEQUENCING: 100,000 reads (full-length cDNAs)
J. M. Cock, Roscoff
- STRAIN SELECTION - BIOLOGICAL MATERIAL - LIBRARIES
- LIBRARIES - SEQUENCING - ASSEMBLY
- AUTOMATIC ANNOTATION - EXPERT ANNOTATION
The Ectocarpus genome (200 Mb) project J.-M. Cock
Cock JM et al. (74 authors) Nature. 2010 Jun 3; 465:617-21 and more than 20 papers during
the two last years.
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
The origin of alginate synthesis route in brown algae is likely the result of an horizontal gene transfer (HGT) from Actinobacteria
Michel et al. New Phytol. 2010 PMID: 20618907
Brown algal genomics
26
Olmix Algae Symposium - Sept. 10th, 2012
Hypothetical biosynthetic pathway for the formation of phloroglucinol derivatives in marine brown algae
(based on Ectocarpus genome data)
Brown algal genomics
27 Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae
27
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Fungi (9) 81
97
88
Bacteroidetes (3) 78
99 Amoebozoa (2)
95
98
Plants (28)
81
100
9898Bacteria (10)
Actinobacteria (7) 100
89 E. siliculosus PKS2
E. siliculosus PKS3 E. siliculosus PKS1
F. vesiculosus PKS F. spiralis PKS S. binderiPKS
76 Brown algae
Chalcone synthases Stilbene synthases
Resveratrol synthases Pyrone synthases Acridone synthase
Bisphenyl synthases …
100
Beta-ketoacyl synthases (outgroup)
Type III Polyketide synthases
Brown algal genomics
28
The origin of phlorotannin in brown algae is likely the result of an horizontal gene transfer (HGT) from Actinobacteria
Phylogenetic relationships of brown algal Type III Polyketide synthases
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium 29
Phloroglucinol synthesis in Pseudomonas fluorescens
Type III Polyketide synthase (PKS)
Boiled control
1 hour
3 hours
5 hours
Abundance
Time →
Extracted ion m/z 342 → Phloroglucinol
CNRS patent sept.2011 n° 11 58728 Meslet et al submitted soon.
Phloroglucinol synthesis in Ectocarpus siliculosus
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Other brown seaweeds
A kelp with major ecological importance
A maricultured kelp
A proposal for a project between Chile, Germany, USA & France submitted to JGI
30
Macrocystis pyrifera (L.) C.Agardh
28
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Red algal genomics
31
The Chondrus genome project
The Porphyra genome projects
Jonas Collén Catherine Boyen
Olmix Algae Symposium - Sept. 10th, 2012
Hana-Tsunomata™ when translated from Japanese means "Flower Chondrus". Other common references to Hana-Tsunomata™ sea vegetables include Hana-nori™ ("Flower seaweed"), Cultivated Chondrus, Kaede-nori ("Maple seaweed") and Hana-sakura-sou ("Cherry blossom sea plant").
Chondrus crispus flowering (NS, Canada, ASP)
An industrial reality
32 Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
The Ulva genome project 150 Mbp
FSU Jena
John Bothwell
Ulva genomics
33
29
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Ulva mariculture
34
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Trends in seaweed blue chemistry
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
IDEALG
36
18 partners: UEB coordination, SBR: UMR 7139-6 teams, UMR 7144-3 teams+ FR2424, CEVA Pleubian, AMURE-UBO, IFREMER Brest, UBS-LBCM Lorient, Agro-Campus, ENSCR, IRISA-Symbiose (Rennes), CNRS U-Nantes, INRA-LBE, 1 large company : Danisco & 4 SMEs, C-Weed, Aleor, France Haliotis, Bezhin Rosko
IDEALG 2011-2020 Seaweed biotechnology and bioresources
A national 10-year large integrative project to merge
algal genomics with mariculture, biotechnology and chemistry.
30
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Knowledge transfer:
37
Apply knowledge of metabolic pathways and networks, enzymes, molecules, biocatalyzers, bioconversion, bioengineering and aquaculture of domesticated seaweeds, Pre-pilot scale projects and technological research
Develop basic research on brown, red and green seaweeds toward domestication of local crops and improvement of seaweeds uses in biotechnology and blue-green chemistry
Domestication
Biotechnology Sustainable Chemistry
Seaweedomics toward
Systems biology
seaweed.ie
Thanks for your attention!
38 G. Michel B. Kloareg
J.M. Cock
M. Czjzek
C. Leblanc
J. Collén
C. Boyen
31
Marine macroalgae: a sweet treasure
Dr. Mirjam Czjzek 1
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Marine seaweeds: a large diversity of original polysaccharides
2 Baldauf (2003) Science
1 cm
Ectocarpus siliculosus
Chondrus crispus
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Marine seaweeds: a large diversity of original polysaccharides
3 Baldauf (2003) Science
fucans
Me
-3OSO
OOSO3-
O
O
OH
Me OOSO3-
Me
-3OSO
OOSO3-
O
O
OH
Me OOSO3-
O
O S O 3 -
O
O H
O
O S O 3 -
O H
O H O
O S O 3 -
agar/carrageenans
O H O
O H
O
O O
O H H O
O
H O H O
O O
H O O H
O H O H
O
xyloglucan
OOO
OH OH
O CH3
OOH
OHO2C
n
X+ -O3SX+ -O3S
ulvan
O
O
OH
OH
O H
O S O 3 -
OH O
O
O
NH
O
dermatan
32
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
The diversity of sugars and their linkages in polymers….
1
4
3 2
5 6
4
1
1 2
….goes hand in hand with the diversity of enzymes
GH GT glycosyl transferases or GT
glycosyl hydrolases or GH
Biosynthesis Catabolism, remodeling
glucomannan
lichenan
cellulose glucose
mannose
galactose
4
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Chlorophyta (Green algae)
Rhodophyta (Red algae)
Heterokonts (Brown algae & diatoms)
Neutral polysaccharides
(mostly shared with land
plants)
Cellulose
Xylan Mannan Chitin
Cellulose
Mannan
Cellulose
Chitin (Diatoms)
Starch Floridean starch Laminarin
Polyanionic polysaccharides
(unique to marine algae)
Macroalgae
Ulvans Agars
Carrageenans Alginates Fucanes
Unicellular algae Capsulans
(Prasinococcus)
Sulfated glucogalactoxylans
(Porphyridium)
Carboxysulfated polysaccharides (Phaeodactylum)
Sulfated carboxylated Carboxy-sulfated
Algal polysaccharides: the complexity of organic matter in ocean
5
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Red algal galactans
agarose
k-carrageenan
i-carrageenan
l-carrageenan
b(1-4) a(1-3) O
O O
O
OH
O
OH OH
OH
O
O OO
OSO3-
O
OH
OH
OH
O
O OO
OSO3-
O
OH
OH
OSO3-
O
OSO3-
O
OH
O
OSO3-
OH
OHO
OSO3-
porphyran
Increase in density of negative charge
Decrease in gel strength
Gels of 1 %
6
33
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Conformational state of k-carrageenan (iono- and thermo-reversible gels)
NaCl NaI KCl
Helices
Variables Temperature [carrageenan]
[salt] Type of salt
Random coils or Flexible rods
Aggregation of helices Gel formation
7
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
b-agarase
Extended loops
k-carrageenase
Structural determinants of the k-carrageenase
8
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Extended loops
k-carrageenase
Structural determinants of the k-carrageenase
9
Trp95 Arg151
34
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
soluble substrate
endo
15 20 25 30 Time (min)
DP4 DP6 DP8
Anion exchange chromatography
µS 0
36
47 49
60
51
10
% hydrolysis
citrate
µS
processive! jellified
substrate
Time (min)
DP4
15 20 25 30
0
18
40
42
56 62
% hydrolysis
10
citrate
0 40 10 20 30 0
0,1
0,2
0,3
Hydrolysis (%)
DP
4/C
itrat
e
Mode of action of the Pseudoalteromonas k-carrageenase
10
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Mode of action of the Pseudoalteromonas k-carrageenase
DP4 DP4 DP4 11
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Agarose sol/gel transition
Electronic Scanner microphotograph of a 2 % agarose gel
Attack of colloidal microgel by Zobellia galactanivorans
12
35
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Agarose sol/gel transition
Handbook of hydrocolloids (Medin, A. S. 1995)
a(1-3)
agarose
b(1-4) O
O
C H 2 O H O H
O O
O
O
O H O H ) ( n
13
Olmix Algae Symposium - Sept. 10th, 2012
Zobellia galactanivorans, (Bacteroidetes) a specialized marine polysaccharide degrader
Barbeyron et al. 2001
114 glycoside hydrolases (GH), 12 Polysaccharide lyases (PL), 17 carbohydrate esterases (CE) and 72 sulfatases !
several polyspecific familes such as 16 GH16
Genome project : counting the carbohydrate active enzymes (CAZymes)
Genome Project – MPI Bremen F. O. Glöckner, M. Bauer, R. Amann
CeBiTech
14
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Marine bacteria secrete specific enzymes for the degradation of algal polysaccharides
15
b-Agarases AgaB
k-Carrageenases
i-Carrageenases
GH96 CBM6 TPS3 CBM6 TPS3 TPS3 TPS3 TPS3 CBM6 a-Agarase
l-Carrageenase
Pseudomonas agaralyticus Flament et al. (2007) Appl Environ Microbiol
Zobellia galactanivorans Jam et al. (2005) Biochem J
AgaA
P. carrageenovora, Z. galactanivorans Barbeyron et al. (1998) Mol Biol Evol
Alteromonas fortis, Z. galactanivorans Barbeyron et al. (2000) J Biol Chem
GH-16 X-70 UNK GH-16
GH-16 Big2 GH-16 CBM16
GH-82 GH-82
WD repeat domain New GH
Pseudoalteromonas carrageenovora Guibet et al (2007) Biochem J
Fucanase
Alginate lyase
GH107 Cad Cad Cad UNK
New PL
Mariniflexile fucanivorans Colin et al. (2006) Glycobiology
Pseudomonas alginovora Chavagnat et al. (1996) Biochem J
AgaB
36
Olmix Algae Symposium - Sept. 10th, 2012
Phylogenetic analysis of Zobellia enzymes from family GH16
16
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
The variable compositions of agaroids
17
Neutral, ideal
Various modifications can be observed like, pyruvate groups, methylation, sulfatation, or even branching .... These modified agar components are often called agaropectins
O
O
CH2OMeOSO3-
O O
O
O
OHOH
O
O
CH2
O O
O
O
OH
OO
CH3HO2C
OH
O
O
CH2OHOH
O O
O
O
OHOH
O
O
CH2OMeOH
O O
O
O
OHOH
O
O
OH
O O
O
O
OHOH
O
HOOH
O
HO
Me
OOMe
CH3HO2COSO3-
a(1-3) b(1-4) b(1-4) a(1-3)
O
HOOH
Agarose
b(1-4) a(1-3)
M. Lahaye and C. Rochas 1991
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Correlation with increasing L-galactose-6-sulfate units Maximum activity on red algae Porphyra sp.
L6S = L-galactose-6-sulfate LA = 3,6-anhydro-L-
galactose
Activity screening on algal cell wall extracts
18
37
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Structural analysis by crystallography
19 Olmix Algae Symposium - Sept. 10th, 2012
W131
L6S L6S
G
G
-4
-3
-2 -1
E144
E139S
W56
H53
R133
R59
catalytic residues
Basic residues are conserved at -2 critical for porphyran recognition
Olmix Algae Symposium - Sept. 10th, 2012
Z. galactanivorans contains a complex agarolytic system
20 Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae
Thanks for your attention!
21
38
Enzymatic hydrolysis in chemistry of seaweeds
Nathalie Bourgougnon, Kevin Hardouin, Loannes Le Bars, Gilles Bedoux, Christel Marty, Justine Dumay*, JP Bergé **
Laboratoire de Biotechnologie et Chimie Marines, UBS, PRES UEB, IUEM * Mer Molécules Santé, Université Nantes ** IFREMER, Nantes 1
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
“Marine biofilm: biological and chemical approaches” program Study of the interactions between organisms and abiotic surfaces Study of physical and chemical parameters involved in bacterial adhesion by
conceiving model substrates Development of antifouling systems combining efficiency and environmental
respect Cell-cell interfaces Complex bacterial biofilms Procaryote-eucaryote interactions and communications
Biotechnology: valorization of marine molecules
Purification and characterization of compounds of interest from invasive marine organisms (sponges, algae, bacteria, echinoderms...)
Laboratoire de Biotechnologie et Chimie Marines
15 teachers/researchers - 10 PhD students - 2 technical assistants
2
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Laboratoire de Biotechnologie et Chimie Marines
Professional Licence Biotechnologie Ingénierie des produits cosmétiques et de santé
Master Biotechnologie Biomolécules, micro-organismes and bio-procédés
3
39
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Which biomasses to screen?
Marine Drug design ?
Extracts with biological activities
Identification of the producer? extraction and purification, Synthesis very difficult, limited resource
Resources halieutics, by-products…?
Few active molecules
Interesting molecules
Many ways of upgrading
Biotechnologic biomass ?
Lot of biological molecules
Simple extraction
Inexhaustible resource
Many ways of upgrading
Selection of phyla know to synthesize cytotoxic molecules
4
Olmix Algae Symposium - Sept. 10th, 2012
French Brittany Seaweed A good biomass
Long tradition of the use of seaweeds
Harvest of marine macroalgae in Brittany (2009)
Brown Red Green Others
94.7%
3.9% 1.1%
0.3%
Important biodiversity
Harvested wild seaweed 70 000T/year CSAVM 2010
Sangiardi 2010
Good economic context with a lot of active companies using macroalgae
Invasive seaweeds
5
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Animal and human foods Health market
Cosmetics
Chemical synthesis of natural substances
Extraction from marine organisms
Technologies
6
40
Olmix Algae Symposium - Sept. 10th, 2012
Extraction from marine organisms
Maceration in a mixture of solvants Liquid/liquid partition Fractionation by chromatography Purification by HPLC Identification RMN, Malditof….
Extraction technologies
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae 7
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Obtain compounds which can be the object of later upgrading from seaweeds
Low energy consummation
Low cost
Processes environment-friendly (no toxic, biodegradable)
Enzymatic hydrolysis
8
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Enzyme molecules
Protein nature Product was not consumed during the reaction Active in small proportion Do not modify the balance thermodynamics reaction Accelerate only the speed of the reaction Specific of a reaction Large-scale production
9
41
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Numbering EC
Enzyme classification
10
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
pH stat method
pH Regulation Temperature Control Calculation of Hydrolysis degree
Temperature Quantity enzyme Time of hydrolysis pH H2O Volume
Parameters studied
Enzymatic hydrolysis technology
11
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Enzymatic hydrolysis technology
12
Raw material (seaweed) (1v)
Enzyme (large spectra) 1.0% Water (1v)
Temperature 60°C
Time 3 hours
Reaction
42
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Example of enzymatic hydrolysis
13
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Leu-Trp-Lys-Arg-Glu-Ile-Tyr-Phe-Arg-Gln-Ser-Val-Asp-Thr-Ala-Pro-Asn
Example of enzymatic hydrolysis
14
Alcalase EC 3.4.21.62
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Leu-Trp-Lys-Arg-Glu-Ile-Tyr-Phe-Arg-Gln-Ser-Val-Asp-Thr-Ala-Pro-Asn
Example of enzymatic hydrolysis
15
Alcalase EC 3.4.21.62
Protamex EC 3.4.21.62 &
EC 3.4.24.28
43
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Leu-Trp-Lys-Arg-Glu-Ile-Tyr-Phe-Arg-Gln-Ser-Val-Asp-Thr-Ala-Pro-Asn
Alcalase EC 3.4.21.62 Flavourzyme
EC 3.4.11.1
Protamex EC 3.4.21.62 &
EC 3.4.24.28
Enzymes tested only or in combination
Example of enzymatic hydrolysis
16
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Enzymatic hydrolysis Process
17
Identification of seaweed biomass Good knowledge of biomass composition
SEAWEEDS
Analysis
Protein Lipid Polymer Mineral
Quality
Food grade Non Food Grade
Quantity
Seasonality Geography …
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Enzymatic hydrolysis Process
18
Define the objectives of upgrading
SEAWEEDS
Analysis
Protein Lipid Polymer Mineral
Quality
Food grade Non Food Grade
44
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Enzymatic hydrolysis Process
19
Define the objectives of upgrading
SEAWEEDS
Analysis
Protein Lipid Polymer Mineral
Quality
Food grade Non Food Grade
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Enzymatic hydrolysis Process
General Strategy 20
Estimate the availability of the biomass
SEAWEEDS
Analysis
Protein Lipid Polymer Mineral
Quality
Food grade Non Food Grade
Quantity
Seasonality Geography …
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Enzymatic hydrolysis Process
21
Analysis
Protein 10-24% Lipid Polymer
38-60% Mineral 14-29%
Quality
Food grade Non Food Grade
Quantity
Seasonality Geography …
g / 100 g MS
45
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Enzymatic hydrolysis Process
22
Total protein by Kjeldhal
Soluble protein by Lowry and Bradford method
0 1 2 3 4 5 6
Temps (min)
Ala
GlyVal
IS
Leu
IleThr
SerPro
Asp
Met
Glu
Phe
Lys Tyr
Liquid chromatography
Gaz chromatography FID
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Enzymatic hydrolysis Process
23
Total sugars by Dubois
Anion exchange chromatography HPAEC
Characterization monosaccharides
Steric exclusion Low pression chromatography
Polysaccharides profil
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Enzymatic hydrolysis Process
24
AQUEOUS
SLUDGE
OILY 15% Proteins
0% Lipids
5% Minerals 24% Proteins
2% Lipids
20% Minerals
23% Sugars
46
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Enzymatic hydrolysis Process
25
Analysis
Protein 10-24% Lipid Polymer
38-60% Mineral 14-29%
Quality
Food grade Non Food Grade
Quantity
Seasonality Geography …
Animal nutrition and animal Health Vegetal health
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Enzymatic hydrolysis Process
26
Studies of seasonality and geographic parameters
Analysis
Protein 10-24% Lipid Polymer
38-60% Mineral 14-29%
Quality
Food grade Non Food Grade
Quantity
Seasonality Geography …
Olmix Algae Symposium - Sept. 10th, 2012
Enzymatic hydrolysis Process
27
Enzyme 2
Enzyme 1
Composés minoritaires
Amadéite®
Protéines, Glucides, Lipides, Minéraux
Oligosaccharides -
Polysaccharides
Protéines
Enzyme 1
Hydrolyse 1
Algues
Culot
Enzyme 2
Hydrolyse 2
Culot
Composés
Minéraux, lipides,…
Nutrition
Autres (non hydrolysable)
Oligosaccharides
Peptides
Protéines
Polysaccharides
47
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Proposed value chains (France) Seaweeds - Recovery of molecules with industrial interest
28 J. Fleurence & J. Dumay (MMS), E. Deslandes &V. Stiger (LEMAR), N. Bourgougnon (LBCM), R. Baron & JP Bergé (Ifremer)
Seaweed biomass
Clarified extract
Solid residues
Seaweed harvesting
Algoculture
SEPARATION CONCENTRATIONPURIFICATION
(membranes or chromatography
processes)
EXTRACTION Enzymatic
digestion, reactive extrusion, or
classical extraction with aqueous solvent in mild
conditions AGRI-FOOD
AQUACULTURE COSMETICS PHARMACY
HEALTH
Enzy
mat
ic tr
eatm
ent
Proteins Bioactive peptides
Pigments Oligosaccharides polysaccahrides
Phenolic compounds
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Proposed value chains (France) Seaweeds - Recovery of molecules with industrial interest
29
Enzymatic extraction of protein from Palmaria palmata Different enzymatic treatment of seaweed
R-Phycoerythrin
Recovery Yield : 4 % expressed in regard of the dry weight instead to 0.4 % with a classical extraction
Thanks for your attention!
30
Young researchers: Kevin Hardouin & Loannes Le Bars Technical assistant: Christel Marty
Collaboration:
48
Bioactivities of Marine Polysaccharides (MSP)
in human and animal health - Uptade
Dr Henri SALMON - Director of research - INRA 37380 Nouzilly Dr Hervé DEMAIS - Scientific advisor - OLMIX 56580 Bréhan
1
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Polysaccharides: Introduction
Polysaccharides = biologically active substances in biochemistry and medicine (aside from their rheological properties).
Today, the most promising biopharmacological activities of polysaccharides are their immunomodulatory and antitumor effects [1].
The most actives being extracted from primitives plants like mushrooms or algae.
2
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Polysaccharides = macroamolecules
Repetitive structural features which are polymers of monosaccharide units joined to each other by glycosidic linkages
3
49
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Classification of the bioactive sulfated polysaccharides
Seaweed
Red Green Brown
Ulvan
Kappa, Iota, Lambda types
Galactan and Carrageenan Fucoidan
Fucose, Xylose,Uronic acid, Galactose, Sulfate
Sulfated agalctose and 3,6 Anhydrogalactose
Sulfated rhamnose, sulfated aldobiuronic
acid
Marine Sulphated polysaccharides (MSP)
4
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Polysaccharides = primary structural variability
Great potential for structural variability due to their interconnections (glycosidic bonds) at several points to form a wide variety of linear or branched structures [2].
Glycogen
α-1,4 ->LINEAR α-1,6 ->BRANCHED
1
2 3
4 5
6
1
1
2 3
4
Amylose
α1,4 glucose-> LINEAR
1 2
3
4
5
6
5
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Marine Sulphated polysaccharides (MSP): ULVANS
6
Sulphated Polysaccharides -> Poly-anionic structure -> various bioactivities
. [→4)-α-l-Idop-(1→4)-α-l-Rham3S-(1→]n
[→4)-β-d-Glcp-(1→4)-α-l-Rham3S-(1→]n
Rare sugar such as Rhamnose (also in plants)
Glucuronic and iduronic acids (sugars from the mammalian glycosaminoglycans
family)
High sulfate content (Sulphated rhamnose)
A
50
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Marine Sulphated polysaccharides (MSP): FUCANS
7
Sulphated Polysaccharides -> Poly-anionic structure -> various bioactivities
1→2, 4-O-sulphated fucopyranose)
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Bioactivities of MSP
Antiviral Activities
Anti-Inflammatory Activities
Immuno Activities Antioxydant Activities
Anticoagulant and Antithrombotic Activities
Antilipidemic Activities
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Antiviral Activities Generalities
Inhibition of the replication of enveloped viruses: blocks the viral entry into the cell by binding to glycoprotein C (gC) and glycoprotein D (gD) of HSV-1(37)
-> Antiviral activity of sulfated polysaccharides increases with sulfation and their molecular weight [34]
Chemical structure of disaccharides (VI and VII) relevant for binding
9
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Antiviral Activities Antiviral Activities
10
In vitro inhibition of influenza A virus infection by marine microalga-derived sulfated polysaccharide p-KG03 Meehyein Kim a, Joung Han Yim , So-Yeon Kim a, Hae Soo Kim ,Woo Ghil Lee a, Sung Jin Kim ,Pil-Sung Kang b, Chong-Kyo Lee
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Immuno-Inflammatory Activities of MSP: Generalities
MSP may affect multiple targets in the immune and inflammatory systems that can have impact on disease progression and outcome including tumor progression and metastasis [41]. Sulfated polysaccharides play two-edged roles, inhibitor and promoter, in immune response.
General Effects : • Stimulation of the immune response / control of immune cell activity to
mitigate associated negative effects such as inflammation [40].
• Anti-inflammatory Growing body of evidence illustrating their ability to interfere with the migration of leukocytes to sites of inflammation. [42,43].
• Inhibit tissue degradative enzymes such as heparanase and elastases that are involved in the breakdown of basement membrane integrity during inflammation [45,46].
11
Olmix Algae Symposium - Sept. 10th, 2012
Intravital microscopic images of a rabbit mesenteric venule. (A)Basal leucocytes rolling along the venular endothelium (B) demonstration that the rolling was abolished 3 min after systemic administration of fucoidin (10 mg/kg).
Changes in basal rolling leucocytes flux without change in arterial blood pressure over time in the rabbit after systemic treatment with fucoidin
By binding of fucoidan to L- and P selectins, cell adhesion molecules essential in the recruitment process., Fucans also inhibit leukocyte recruitment to the abdominal cavity during acute peritonitis in rats [44].
Anti-Inflammatory Activities of MSP: Anti-migration of leukocytes to sites of inflammation
12
interfering with the migration of leukocytes to sites of inflammation. [42,43].
52
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Anti-Inflammatory Activities of MSP By Anti-Complement activity
Fucoidan fractions (low MW) inhibit both the classical and alternative pathways in human serum [47] by binding to the C1q and C4 of the complex
-> interaction with the complement ->reduce the pro-inflammatory state
X D, B, F :Area of
interaction with C4
13
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
λ-carrageenan stimulate mouse T cell cultures in a toll-like receptor-4 (TLR4) dependent manner generating a T helper 1 (Th1) patterned cytokine response. However, splenocytes prepared from TLR4-deficient mice still retain some ability to produce interferon-γ in response to λ-carrageenan suggesting that PRRs other than TLR4 are also elicited. [52]
[40].
Immuno-Activities of MSP: Binding to PRR
Pattern Recognition Receptors (PRRs)
14
mannose Toll-Like Receptors
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
These and other reports of algal sulfated polysaccharides directly stimulating the innate immune system [53,56,57] suggests that they may find therapeutic use in opposing T helper 2 (Th2)-based pathologies such as autoimmune disorders and allergy.
(56) (56)
Glial cells + TNFα +IFN=>iNOS (inducible nitric oxide synthase )
Inbition iNos with doses of Fucoidan
TNFα +IFN
TNFα +IFN TNFα +IFN+IFNTNFα +IFN+IFNFucoidan (µg/ml)
Fucoidan(µg/ml)
Anti-Inflammatory properties of MSP: Inhibition of iNOS production
15
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Immuno- Activities of MSP: Via Macrophage activation
16
Botanical polysaccharides: Macrophage immunomodulation and therapeutic potential Igor A. Schepetkin, Mark T. Quinn *
Fucoidan+DC A pro-inflammatory cytokine/chemokine profile [53, 54, 55,56].
Chemokines
MCP-1
MSP
ROS=reactive oxygen production
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
57. Kim, M.H.; Joo, H.G. Immunostimulatory effects of fucoidan on bone marrow-derived dendritic cells. Immunol Lett 2008, 115, 138-143.
58. Choi, E.M.; Kim, A.J.; Kim, Y.O.; Hwang, J.K. Immunomodulating activity of arabinogalactan and focoidan in vitro. J Med Food 2005, 8, 446-453.
Fucoidans, carrageenan
tumoricidal activity
Anti-tumoral activities of MSP
17
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Immuno-Inflammatory Activities of MSP: Summary of immune cells Activation by MSP
18
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
MSP: Anticoagulant Activity
MSP (Fucoidans, and ulvan)
and Red seaweeds >Heparin-like anticoagulant activity
[8,9,10 ,11,12]. [13,14,15,16,17].
Some high rhamnose-containing MSP are
more potent than standard heparin [16]. Control: Clotting time: 40s
Anti-thrombic 19
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
MSP: Antioxidant Activities
Ferric reducing/antioxidant power [23]
Superoxide radical scavenging ability [24,25, 26, 22].
Fucans’ Superoxide radical scavenging activity correlated positively high sulfate content of the polysaccharide fractions [24,26]. Antioxidant properties of Carrageenans [24] and Ulvans [27 ] also appeared related to sulfate content.
Inhibition of lipid peroxydation of rat liver microsome
20
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
MSP: Antilipidemic Activities
21
Lipid-lowering (serum triglyceride and total cholesterol) in hyperlipidemic animal models (28,29,30).
In rats fed a high cholesterol diet for 21 days, supplementation of the diet with ulvans from U. pertusa led to reductions in serum total cholesterol and LDL-cholesterol with no significant alteration in serum triglycerides . The effects of ulvans were modified when it was degraded into lower molecular weight fractions (29).
Illustration of the function of hepatic lipase as a lipolytic enzyme. Santamarina-Fojo S et al. Arterioscler Thromb Vasc Biol 2004;24:1750-1754
M.S.P
Certainly fucoidan and other algal sulfated polysaccharides may influence LPL (Lipoprotein Lipase) and HL (Hepatic Lipase) through interaction with well-characterized heparin-binding sites on these enzymes (31).
Conclusion:Algal sulfated polysaccharides are showing promising effects in addressing the hyperlipidemia associated with certain drug toxicities (32, 33).
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Conclusions 1
The study of algal sulfated polysaccharides structures is challenging, because of their diversity and heterogeneity.
This may also have hindered their development as therapeutic agents to date in spite of various biological activities already demonstrated. (Immunity, inflammation, coagulation, oxidation, lipidemia….)
Due to the difficulties in identifying the precise chemical structure of algal sulfated polysaccharides, the relation between their structures and biological activities is far from fully understood.
The production of a standardized commercial product based on algal sulfated polysaccharide constituents can also be a challenge since their structural and pharmacological features may vary depending on species and on location and time of harvest [66].
22
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Conclusions 2
23
Important points can already be mentioned regarding the conditions of MSP bioactivities:
Importance of understanding the structural requirements for biological activity
Do low molecular weight derivatives, which are potentially more bioavailable, remain active?
Therapeutic use by oral route can be limited due to low bioavailability given often high molecular weights of MSP(64).
At the opposite :
Advantage for the hypolipidemic effects due to bile acid sequestration in the intestinal lumen.
With regards to some immunomodulatory activities, site of activation of the immune system may also be within the intestinal lumen (e.g., at Peyer’s patches).
Hypothesized for immunomodulatory effects of polysaccharide constituents from Chlorella pyrenoidosa [65].
Olmix Algae Symposium - Sept. 10th, 2012
Conclusions 3
= increased activity = reduced activity = increased activity = reduced activity= increased activity = reduced activity= increased activity = reduced activity
Algal sulfated polysaccharides are a new source of numerous biological activities that may find in human and animal health many prophylactic and
therapeutic benefits in the near future.
24
56
Thanks for your attention!
25
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
6. Lahaye, M.; Robic, A. Structure and functional properties of ulvan, a polysaccharide from green seaweeds. Biomacromolecules 2007, 8, 1765-1774.
5. Percival, E.; McDowell, R.H. Chemistry and Enzymology of Marine Algal Polysaccharides; Academic Press: New York, NY, USA, 1967; p. 219.
7. Lahaye, M.; Brunel, M.; Bonnin, E. Fine chemical structure analysis of oligosaccharides produced by an ulvan-lyase degradation of the water-soluble cell-wall polysaccharides from Ulva sp. (Ulvales, Chlorophyta). Carbohydr Res 1997, 304, 325-333.
2. Sharon, N., Lis, H. Scientific American, 1993, pp. 74-81.
3. Hodgson, J. Biotechnology, 1991, 9, 609-613.
1. Ooi, V.E.C; Liu, F. Immunomodulation and Anti-Cancer Activity of Polysaccharide-Protein Complexes. Current Medicinal Chemistry, 2000, 7, 715-729.
4. Lahaye, M.; Ray, B. Cell-wall polysaccharides from the marine green alga Ulva rigida (Ulvales, Chlorophyta)-NMR analysis of ulvan oligosaccharides. Carbohydr Res 1996, 283,161-173.
8. Bernardi, G.; Springer, G.F. Properties of highly purified fucan. J Biol Chem 1962, 237, 75-80.
9. Springer, G.F.; Wurzel, H.A.; McNeal, G.M.; Ansell, N.J.; Doughty, M.F. Isolation of anticoagulant fractions from crude fucoidin. Proc Soc Exp Biol Med 1957, 94, 404-409.
10. Kusaykin, M.; Bakunina, I.; Sova, V.; Ermakova, S.; Kuznetsova, T.; Besednova, N.; Zaporozhets, T.; Zvyagintseva, T. Structure, biological activity, and enzymatic transformation of fucoidans from the brown seaweeds. Biotechnol J 2008, 3, 904-915.
11. Li, B.; Lu, F.; Wei, X.; Zhao, R. Fucoidan: structure and bioactivity. Molecules 2008, 13,1671-1695.
12. Pomin, V.H.; Mourao, P.A.S. Structure, biology, evolution, and medical importance of sulfated fucans and galactans. Glycobiology 2008, 18, 1016-1027.
13. Matsubara, K.; Matsuura, Y.; Bacic, A.; Liao, M.L.; Hori, K.; Miyazawa, K. Anticoagulant properties of a sulfated galactan preparation from a marine green alga, Codium cylindricum. Int J Biol Macromol 2001, 28, 395-399.
14. Farias, E.H.C.; Pomin, V.H.; Valente, A.P.; Nader, H.B.; Rocha, H.A.O.; Mourao, P.A.S. A preponderantly 4-sulfated, 3-linked galactan from the green alga Codium isthmocladum.Glycobiology 2008, 18, 250-259.
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18. Farias, W.R.L.; Valente, A.P.; Pereira, M.S.; Mourao, P.A.S. Structure and anticoagulant activity of sulfated galactans. Isolation of a unique sulfated galactan from the red algaeBotryocladia occidentalis and comparison of its anticoagulant action with that of sulfated galactans from invertebrates. J Biol Chem 2000, 275, 29299-29307.
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35. Luescher-Mattli, M. Algae, A Possible Source for New Drugs in the Treatment of HIV and Other Viral Diseases. Curr Med Chem 2003, 2, 219-225.
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31. Yokota, T.; Nagashima, M.; Ghazizadeh, M.; Kawanami, O. Increased effect of fucoidan on lipoprotein lipase secretion in adipocytes. Life Sci 2009, 84, 523-529.
32. Raghavendran, H.R.; Sathivel, A.; Devaki, T. Effect of Sargassum polycystum(Phaeophyceae)-sulphated polysaccharide extract against acetaminophen-induced hyperlipidemia during toxic hepatitis in experimental rats. Mol Cell Biochem 2005, 276, 89-96.
33. Josephine, A.; Veena, C.K.; Amudha, G.; Preetha, S.P.; Varalakshmi, P. Protective role of sulphated polysaccharides in abating the hyperlipidemic nephropathy provoked by cyclosporine A. Arch Toxicol 2007, 81, 371-379..
25. Zhao, X.; Xue, C.; Cai, Y.; Wang, D.; Fang, Y. Study of antioxidant activities of fucoidan from Laminaria japonica. High Tech Lett 2005, 11, 91-94.
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27. Qi, H.; Zhang, Q.; Zhao, T.; Chen, R.; Zhang, H.; Niu, X.; Li, Z. Antioxidant activity of different sulfate content derivatives of polysaccharide extracted from Ulva pertusa(Chlorophyta) in vitro. Int J Biol Macromol 2005, 37, 195-199.
34. Witvrouw, M.; De Clercq, E. Sulfated polysaccharides extracted from sea algae as potential antiviral drugs. Gen Pharmacol 1997, 29, 497-511.
37. Harden, E.A.; Falshaw, R.; Carnachan, S.M.; Kern, E.R.; Prichard, M.N. Virucidal activity of polysaccharide extracts from four algal species against herpes simplex virus. Antiviral Res2009, 83, 282-289.
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47. Blondin, C.; Fischer, E.; Boisson-Vidal, C.; Kazatchkine, M.D.; Jozefonvicz, J. Inhibition of complement activation by natural sulfated polysaccharides (fucans) from brown seaweed.Mol Immunol 1994, 31, 247-253.
41. Groth, I.; Grunewald, N.; Alban, S. Pharmacological profiles of animal- and nonanimal-derived sulfated polysaccharides--comparison of unfractionated heparin, the semisynthetic glucan sulfate PS3, and the sulfated polysaccharide fraction isolated from Delesseria sanguinea. Glycobiology 2009, 19, 408-417.
42. Granert, C.; Raud, J.; Xie, X.; Lindquist, L.; Lindbom, L. Inhibition of leukocyte rolling with polysaccharide fucoidin prevents pleocytosis in experimental meningitis in the rabbit. J Clin Invest 1994, 93, 929-936.
39. Carlucci, M.J.; Scolaro, L.A.; Noseda, M.D.; Cerezo, A.S.; Damonte, E.B. Protective effect of a natural carrageenan on genital herpes simplex virus infection in mice. Antiviral Res2004, 64, 137-141.
43. Preobrazhenskaya, M.E.; Berman, A.E.; Mikhailov, V.I.; Ushakova, N.A.; Mazurov, A.V.; Semenov, A.V.; Usov, A.I.; Nifant’ev, N.E.; Bovin, N.V. Fucoidan inhibits leukocyte recruitment in a model peritoneal inflammation in rat and blocks interaction of P-selectin with its carbohydrate ligand. Biochem Mol Biol Int 1997, 43, 443-451.
45. Senni, K.; Gueniche, F.; Foucault-Bertaud, A.; Igondjo-Tchen, S.; Fioretti, F.; Colliec-Jouault, S.; Durand, P.; Guezennec, J.; Godeau, G.; Letourneur, D. Fucoidan a sulfated polysaccharide from brown algae is a potent modulator of connective tissue proteolysis.Arch Biochem Biophys 2006, 445, 56-64.
46. Parish, C.R.; Freeman, C.; Hulett, M.D. Heparanase: a key enzyme involved in cell invasion. Biochim Biophys Acta 2001, 1471, M99-M108.
44. Cumashi, A.; Ushakova, N.A.; Preobrazhenskaya, M.E.; D’Incecco, A.; Piccoli, A.; Totani, L.; Tinari, N.; Morozevich, G.E.; Berman, A.E.; Bilan, M.I.; Usov, A.I.; Ustyuzhanina, N.E.; Grachev, A.A.; Sanderson, C.J.; Kelly, M.; Rabinovich, G.A.; Iacobelli, S.; Nifantiev, N.E. A comparative study of the anti-inflammatory, anticoagulant, antiangiogenic, and antiadhesive activities of nine different fucoidans from brown seaweeds. Glycobiology 2007, 17, 541-552.
38. Damonte, E.B.; Matulewicz, M.C.; Cerezo, A.S. Sulfated seaweed polysaccharides as antiviral agents. Curr Med Chem 2004, 11, 2399-2419.
48. Clement, M.J.; Tissot, B.; Chevolot, L.; Adjadj, E.; Du, Y.; Curmi, P.A.; Daniel, R. NMR characterization and molecular modeling of fucoidan showing the importance of oligosaccharide branching in its anticomplementary activity. Glycobiology 2010, 20, 883-894.
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53. Leiro, J.M.; Castro, R.; Arranz, J.A.; Lamas, J. Immunomodulating activities of acidic sulphated polysaccharides obtained from the seaweed Ulva rigida C. Agardh. Int Immunopharmacol 2007, 7, 879-888.
49. Tissot, B.; Montdargent, B.; Chevolot, L.; Varenne, A.; Descroix, S.; Gareil, P.; Daniel, R. Interaction of fucoidan with the proteins of the complement classical pathway. Biochim Biophys Acta 2003, 1651, 5-16.
50. Tissot, B.; Daniel, R. Biological properties of sulfated fucans: The potent inhibiting activity of algal fucoidan against the human complement system. Glycobiology 2003, 13, 29G-31G.
51. Tissot, B.; Gonnet, F.; Iborra, A.; Berthou, C.; Thielens, N.; Arlaud, G.J.; Daniel, R. Mass spectrometry analysis of the oligomeric C1q protein reveals the B chain as the target of trypsin cleavage and interaction with fucoidan. Biochemistry 2005, 44, 2602-2609.
52. Tsuji, R.F.; Hoshino, K.; Noro, Y.; Tsuji, N.M.; Kurokawa, T.; Masuda, T.; Akira, S.; Nowak, B. Suppression of allergic reaction by lambda-carrageenan: toll-like receptor 4/MyD88-dependent and -independent modulation of immunity. Clin Exp Allergy 2003, 33, 249-258.
54. Nakamura, T.; Suzuki, H.; Wada, Y.; Kodama, T.; Doi, T. Fucoidan induces nitric oxide production via p38 mitogen-activated protein kinase and NF-kB-dependent signaling pathways through macrophage scavenger receptors. Biochem Biophys Res Commun2006, 343, 286-294.
55. Yang, J.W.; Yoon, S.Y.; Oh, S.J.; Kim, S.K.; Kang, K.W. Bifunctional effects of fucoidan on the expression of inducible nitric oxide synthase. Biochem Biophys Res Commun 2006,346, 345-350.
56. Do, H.; Pyo, S.; Sohn, E.H. Suppression of iNOS expression by fucoidan is mediated by regulation of p38 MAPK, JAK/STAT, AP-1 and IRF-1, and depends on up-regulation of scavenger receptor B1 expression in TNF-alpha- and IFN-gamma-stimulated C6 glioma cells. J Nutr Biochem 2010, 21, 671.
57. Kim, M.H.; Joo, H.G. Immunostimulatory effects of fucoidan on bone marrow-derived dendritic cells. Immunol Lett 2008, 115, 138-143.
58. Choi, E.M.; Kim, A.J.; Kim, Y.O.; Hwang, J.K. Immunomodulating activity of arabinogalactan and focoidan in vitro. J Med Food 2005, 8, 446-453.
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61. Nishino, T.; Yokoyama, G.; Dobashi, K.; Fujihara, M.; Nagumo, T. Isolation, purification, and characterization of fucose-containing sulfated polysaccharides from the brown seaweed Ecklonia kurome and their blood-anticoagulant activities. Carbohydr Res 1989,186, 119-129.
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63. Pomin, V.H.; Pereira, M.S.; Valente, A.P.; Tollefsen, D.M.; Pavao, M.S.G.; Mourao, P.A.S. Selective cleavage and anticoagulant activity of a sulfated fucan: Stereospecific removal of a 2-sulfate ester from the polysaccharide by mild acid hydrolysis, preparation of oligosaccharides, and heparin cofactor II-dependent anticoagulant activity. Glycobiology2005, 15, 369-381.
66. Bourgougnon, N.; Lahaye, M.; Quemener, B.; Chermann, J.C.; Rimbert, M.; Cormaci, M.; Furnari, G.; Kornprobst, J.M. Annual variation in composition and in vitro anti-HIV-1 activity of the sulfated glucuronogalactan from Schizymenia dubyi (Rhodophyta, Gigartinales). J Appl Phycol 1996, 8, 155-161.
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Marine lipids in amplifying cancers chemotherapy
Pr. Philippe Bougnoux - INSERM U 1069, Centre Henry S. Kaplan, University Hospital, Tours, France
philippe.bougnoux@inserm.fr 1
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Background
Tumor sensitivity to anticancer agents is highly variable
Associated with specific breast cancer pathological types (luminal type, basal-like type etc..)
Some of the variability is accounted for by the type of acquired somatic genetic alterations
or tumor-host interactions
or even the host-environment interactions
The prediction of tumor sensitivity to anticancer drugs remains uncertain
There must be dietary determinants of tumor chemosensitivity
How to identify potential dietary components associated ? 2
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Overview
How was DHA identifed: an observational study in breast cancer DHA increases the sensitivity of breast cancer cell lines to anthracyclines Dietary DHA makes rat mammary tumors sensitive to anthracyclines DHA increases radiosensitivity of mammary tumors in rats
How to account for the tumor specificity of DHA action ? Loss of antioxidant defences during tumor progression Remodeling of tumor neovascular architecture
Translation: from the sea to the patient A phase I-II study of DHA supplementation in metastatic breast cancer during
chemotherapy In progress:... a randomized, phase III study
3
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
How was DHA identified as a lipid component of diet associated with tumor chemosensitivity ? An observational clinical study
Context: Breast cancer patients presenting with a tumor larger than 3 cm
receive neoadjuvant chemotherapy as their initial treatment
Some tumors will shrink, others will not
Hypothesis: If dietary lipids influence breast tumor sensitivity to chemotherapy, then patients with sensitive tumors should have past dietary intake of lipids different from that of patients with resistant tumors
4
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
The Adipose Tissue
Precise measure of dietary lipid intake difficult to establish
FA composition reflects past dietary intake of FA Low turnover : not influenced by the last meal
Use of adipose tissue fatty acid profile as a biomarker of past dietary intake of fatty acids
5
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
56 patients with locally advanced breast cancer (or T > 3 cm)
Adipose tissue fatty acid composition on biopsy
Neo-adjuvant chemotherapy (anthracyclins, cyclophosphamide, 5 FU)
Endpoint: Tumor response to neoadjuvant chemotherapy
DHA content of mammary adipose tissue is an independent predictive factor of tumor
chemosensitivity
0
1
2
3
4
5
median >
Odd
s R
atio
*
Adipose tissue DHA, (%)
15
20
Likelihood of response to chemotherapy *
*Adjusted for age, body mass index and tumour size; p=0.03
Bougnoux et al, Br J Cancer, 1999
Fatty acid profile
Study design
6
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
… Could DHA increase the sensitivity of breast cancer cell lines to anticancer
agents ?
DHA is associated with a greater efficacy of chemotherapy…
…causality ?
DHA, docosahexaenoic acid is a 22C, 6 double bonds PUFA In the diet, it is exclusively found in sea products
7
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
DHA enhances doxorubicin efficacy The effect is increased by prooxidant agents
and abolished by antioxidants Germain et al., Int J Cancer, 1998
Control Docosahexaenoic acid (DHA)
Prooxidants Antioxidants
Oleic acid
0 20 40 60 80 100
Cell Viability (%) Conditions
Doxo + DHA + Antioxidants Doxo+ oleic acid +Oxidants
Doxo + Oxidants *
* Doxo + DHA Doxo + DHA + Oxidants
Doxorubicin, 10-7 M (Doxo)
Culture medium enriched (5d) in DHA or oleic acid
Breast cancer cell line MDA-MB 231
Effect of DHA on chemosensitivity of MDA-MB 231
8
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Fatty acids + oxidants
*
*
* *
*
Fatty acids + antioxidants
-10
10
30
50
70
90
% change in cell toxicity after doxorubicin
Fatty acids alone
*
*
*
Fatty acids , 10µg/ml
LA ALN n-3
AA n-6
EPA n-3
GLA n-6
DHA n-3 n-6
R 2 = 0,8293
0
20
40
60
80
100
0 10 20 30 40
hydroperoxides, pmol/µg proteins
Cyt
otox
icity
, %
… while producing lipoperoxides
PUFA enhance cytotoxic efficacy of doxorubicin…
Germain et al., Int J Cancer,1998 Doxorubicine is a quinone macromolecule
which generates an oxidative stress
What about other PUFA?
9
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
In vivo?
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Olmix Algae Symposium - Sept. 10th, 2012
Dietary DHA is readily incorporated into adipose tissue
11 Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium Olmix AlgaeOlmix Algae Symposium
In-vivo dietary intervention with DHA
0
1
2
3
4 5
6
7 8
0.1
0.8
1.8
5.4
Palm DHA 0.1 g
DHA 0.3 g
DHA 0.8 g
Diet
Adipose tissue
Basal diet (7 % peanut/rapeseed) • Control (addition of 8 % palm oil) • DHA (addition of 8 % DHASCO)
•w or w/o antioxidants •w or w/o oxidants
Study design
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Weeks
Cha
nge
in s
ize
of
Targ
et tu
mor
, %
Bougnoux et al, Lipids, 1999
Dietary DHA enhances the antitumor action of anthracyclins
DHA + oxidants
DHA
DHA + anti-oxidants
1 3 5
-40
-20
0
20
40
Palm oil
Epirubicin injections
The oxidative status of diet strongly influences the effect
Epirubicin efficacy according to dietary supplementation
12
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Dietary DHA increases the sensitivity of mammary tumors to ionizing radiation
This effect is abolished by Vit. E
Tumor irradiation
Tum
or s
urfa
ce v
aria
tion,
(%)
-70 -60 -50 -40 -30 -20 -10
0 10
0 2 4 6 8 10 12 14 16 18
Days following irradiation
Palm oil
Diet :
Single dose, 18 Gy
DHA
Colas et al, Int J Cancer 2004
What about other anticancer treatments which generate ROS, such as radiation therapy ?
13
Study design • Dietary enrichment of rat tissues with DHA • Irradiation of mammary tumor with a single
dose of 18Gy • Measure of tumor regression according to diet
Context: Interaction of ionizing radiation with tissues H20 generates reactive oxigen species
Hypothesis: Dietary enrichment of rat tissues with DHA should make them more susceptible to lipid peroxidation induced by ROS
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium 14
What about non tumor tissues ?
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
20
100
200mmHg
20
100
200mmHg
25mm/sec
60 50
40
30 20
10 0
LVDP (mmHg) Basal
+ pro-oxidant contrôle +anti-oxidant
60 50
40 30 20 10 0
LVDP (mmHg) After volume loading
+ pro-oxidant contrôle Diet with 15% fish oil
+anti-oxidant
Germain et al., Pharmacol. Res, 2003
Dietary omega-3 PUFA do not increase the cardiac toxicity of anthracyclins
Saline
ECG
Volume charge
Pressure transductor
Investigation of cardiac toxicity
15
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Loss of antioxidant defences during tumor progression may account for the tumor specificity of DHA action
Maheo et al., Free Rad Biol & Med, 2004
Vibet et al., Free Rad Biol & Med, 2008
How to account for the tumor specificity of DHA action ?
1. Using two breast cancer cell lines at different degree of tumor progression, we found that the degree of tumor progression influenced tumor response amplification by DHA
2. DHA enhanced doxorubicine toxicity only in cell lines able to produce ROS in response to doxorubicine ….and to subsequently produce lipoperoxides
3. In these cell lines, there was a lack of GPx response to the oxidative stress
4. There was an inverse correlation between GPx1 activity and response of the cells to chemotherapy
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Mechanisms other than ROS-induced lipid peroxidation?
… Is the antitumor action of docetaxel amplified by DHA ?
Docetaxel is a macrocyclic molecule without semi-quinone functions
17
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
MDA-MB-231
0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1.00
20
40
60
80
100 ControlDHAControl +Vit EDHA+VitE
Docetaxel nM
Cel
l via
bilit
y, %
… and the sensitizing effect of DHA to docetaxel
is not abolished by vit E
Breast cancer cell line MDA-MB 231
U 921 Tours – Unpublished data 18
Dietary DHA increases the antitumor action
of docetaxel
Chemotherapy with docetaxel Tumor Surface
- 4 - 3 - 2 - 1 0 1 2 3 4 5 6 7
- 70
- 60
- 50
- 40
- 30
- 20
- 10
0
10
20
Control
weeks
Docetaxel
varia
ti on
%
DHA
- 4 - 3 - 2 - 1 0 1 2 3 4 5 6 7
- 70
- 60
- 50
- 40
- 30
- 20
- 10
0
S. Vibet, PhD Theses Tours
Mammary tumors in rats
DHA sensitizes breast cancer cells to docetaxel …
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
DHA increases the activity of several drugs with distinct modes of action
19
The cancer cell Lipoperoxidation through amplified oxidative stress (Vibet et al, 2008)
Anthracyclins: doxorubicin, epirubicin (Germain et al, 1998, Hardman et al, 2001) Ionizing Radiations (Colas & Bougnoux, 2004)
Tubulin kinetics ? Vincristine (Ikushima et al, 1991), Vinorelbin (Menendez et al 2004),
NF-kappa B pathway ? Taxanes (Shaikh & Wahle, 2008) Restores apoptotic pathways ?
The host: Activation of reducing enzymes : Mitomycin C (Pardini et al, 1993) Alteration of drug metabolism through CYPs ..
Cyclophosphamide (Shao & Pardini , 1995)
The tumor microenvironment Generates secondary products with cytotoxic activities ? Changes the tumor microenvironment
- alters immune response against tumor cells (Calder et al, 2009) - alters angiogenesis, through lipid peroxidation ?
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Investigating tumor neovascular architecture
20
Using Power-Doppler sonography of rat mammary tumors, sensitized with microbubles to quantify tumor vessels We found that dietary DHA led to decreased tumor vascularization, prior to any chemotherapy, and that effect of DHA was abolished by dietary Vit. E (Colas et al., Clin Cancer Res 2006)
Remodelling tumor neovessels may also account for the tumor specificity of DHA action
Denis et al., Clin Cancer Res 2003
Using polymer casts of tumor blood vessels, we found that fish oil diet induced remodeling of tumor vascularization with lower density and thiner blood vessels This led to decreased interstitial pressure within mammary tumors and increased diffusion
Goupille et al, Breast Cancer Res & Treat 2012
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Translation : A pilot study of chemosensitization In metastatic breast cancer patients
Need for a proof of concept
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Evaluation of the feasibility and safety of DHA administration during chemotherapy of metastatic breast cancer
Hypothesis: 1.8 g of daily DHA should lead to an enrichment of membrane lipids of breast cancer metastases, making them more sensitive to anthracyclin-based chemotherapy
Study design: Phase I-II study, with 25 patients – monocentric (Tours) – 2 years inclusion - Patients with metastatic breast carcinoma, OMS < 2 - DHASCO (Martek Inc) 9 cp/day, 10 days prior to chemotherapy, up to the end of CT - Chemotherapy with Epirubicine, Cyclophosphamide, 5-FU, 1 cycle every 3 weeks
End points: Safety & compliance, plasma + RBC levels of DHA Time to tumor progression
Secondary end points: Survival Bougnoux et al., Proc Am Ass Cancer Res, 2006
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Interindividual variability in DHA incorporation after dietary intervention
Stratification into 2 groups
High DHA incorporators
Low DHA incorporators
DHA is rapidly incorporated after dietary intervention
Plasma level
Red Blood Cells
Results:
23 Olmix Algae Symposium
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Elevated DHA incorporation during first line chemotherapy improves survival
Bougnoux et al, Br J Cancer 2009
Median follow-up = 18.6 months (range 3 to 53 months)
p<0.007 p<0.02
Survival according to the incorporation status of DHA
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Vascularization ? The example of liver metastases
25 Olmix Algae Symposium - Sept. 10th, 2012
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Discussion
Increasing plasma DHA level results in clinical benefit in patients undergoing chemotherapy for metastatic breast cancer
Supplementation with DHA is safe, and may reduce some side effects
The variability in DHA incorporation among patients may be circumvented by providing DHA within a food carrier and not as capsules of oil
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
No, this is premature PUFA should not be provided without defined clinical trials
A phase III randomized trial comparing the fatty acid to placebo is on-going in metastatic breast cancer patients to definitely assess the usefulness of fatty acid supplementation during chemotherapy
Finally, could dietary DHA or n-3 PUFA be provided safely during cancer treatments ?
Not with antioxidant molecules at pharmacological doses …may stimulate tumor growth
Effect may change according to the intake of other PUFA ?
EPA has been reported to alter immune response and promote melanoma growth in experimental systems
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
RAN
DO
MIZ
E
(Vegetal oil in food supplement)
Fish oil, 2 g / day
First line chemotherapy (anthracyclines, taxanes..)
Metastatic breast cancer
HER2 negative
RH positive
(Fish oil in food supplement) DHA 1.5 g / day
DHA 0 g / day
216 patients, 2 yrs inclusion
Dietary intervention, 4 to 6 months
Endpoints: Time to progression QoL Funding : PHRC 2011
15 cancer centers involved
The future… marine-derived lipids as adjuvant to breast cancer treatments Bougnoux et al, Prog Lipid Res 2010
ClinicalTrials.gov Identifier: NCT01548534
Aim of the study: To determine whether adjuvant nutrition improves the treatment of breast cancer (Dec 2011 - July 2015)
A randomized, double-blind, phase III multicentric clinical trial comparing fish oil supplementation versus vegetable oil during chemotherapy in metastatic breast cancer patients
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Supports: Région Centre, French Research Ministry , DHOS, Inserm (ATC), INCa (Canceropôles)
Maastricht
• NICE
Participants
Inserm U 1069, Clinical oncology CHU Tours, France Charles Couet Caroline Goupille Marie-Lise Jourdan Emmanuelle Germain Séverine Colas Karine Mahéo Nawale Hajjaji
Oncology & Radiotherapy Olivier Le Floch Agnès Reynaud-Bougnoux
Surgical Gynecology Gilles Body Jacques Lansac
29
CIC Inserm 202, Tours Bruno Giraudeau
Tumor imaging Léandre Pourcelot François Tranquart Inserm U 930 Tours
Phase III randomized clinical trial Virginie Berger + CECO ICO-CPP, Angers France
Thanks for your attention!
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CHILE Reliable Supplier of Quality Food
Eliana Henríquez Flores, Ing. Agrónomo Information Center of Natural Resources
Head of International Affairs Unit 1
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Exceptional sanitary conditions: Phytosanitary Island
Southern hemisphere off-season agricultural production
Chile, long and narrow country, located in South America between latitudes 17 º 30 'and 56 º 30' south latitude.
Diversified Production: More than 5 000 km long, with 16 of the 24 climates present in the world.
5.1 million ha. arable land in an area of 75 million ha.
Population: 17,248,450 (13% rural)
Diversity of climates: Diversity production North: Desert West: Pacific Ocean East: Andes mountains South: Southern Ice
Chile
2
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Chile
3
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
MINISTRY OF AGRICULTURE
Oficina de Estudios y Políticas Agrarias Servicio
Agrícola y Ganadero
Comisión Nacional de
Riego
Instituto de Desarrollo
Agropecuario
Corporación Nacional Forestal
Instituto de Investigaciones Agropecuarias
Fundación Comunicación Capacitación y Cultura del
Agro
Information
Center of Natural
Resources
Fundación para la
Innovación Agraria
Instituto Forestal
Agencia Chilena para la Calidad e Inocuidad
Alimentaria
Institutions of the Ministry of Agriculture
4
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Information Center of Natural Resources
It is an institution that provides information on renewable natural resources, which has brought together the largest database of georeferenced soil, farms, water resources, agroclimate, fruit plantations and forest that exists in Chile, and other products are developed for different MINAGRI institutions.
5
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
General scenario of algae in Chile
In Chile, seaweeds are exported as raw material, used internally as alginates and agar and to a lesser degree, are also consumed as food.
The species that accumulate polysaccharides in their walls are a source of raw material for a variety of industrial products which the agar, carrageenan and alginates are the most used.
Seaweed synthesized polysaccharides in considerable amounts constituting an important renewable resource. A limited number of red and brown algae are exploited for the production of industrial application phycocolloids. It has been found that some seaweed sulfated polysaccharides have interesting biological properties such as anticoagulant activity, antitumor and antiviral.
The production of algae (gracilarias) borders the 90 000 t per year and there are only emerging projects that seek to increase diversity in the cultivation of these plants hydrobiological.
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SPECIE DESCRIPTION
Macrocrystis pyrifera The common name of this alga is "huiro" or "kelp". The contribution of this alga is in the production of alginates. In Chile extends from Cape Horn to Valparaiso. (SANTELICES, 1989).
Porphyra columbina Conocida como "luche rojo". Es explotado casi de toda la costa Chilena, comercializándose a nivel local para el consumo humano bajo la forma de precosido y seco. (RED ALGAS MARINAS CHILE, 1990).
Lessonia trabeculata Known as "red fight." Is exploited almost all the Chilean coast, locally marketed for human consumption in the form of pre-cooked and dry. (RED SEAWEED CHILE, 1990).
Ulva lactuca Its common name is "fight", "fight green" and "sea lettuce". One of the most common species off the coast of the Tenth Region of Chile. Consumption in the region is low, its main contribution of carbohydrates. (Ramirez, 1981.).
Gracilaria chilensis
Common name "nap". The main raw material used in production of Agar-Agar is unique to the genre Glacilaria. (SANTELICES, 1986). The main importing country of Chile is Japan, which absorbs about 80% of production. Coquimbo is the main area of export of this resource. (SANTELICES, 1990).
Durvillaea antartica
In our country known as cochayuyo. In Chile this plant is harvested for human consumption or for export as raw material for the production of alginates. Human consumption takes two forms. The conical stipe is consumed fresh, as part of salads or stews and is called "ulte".Fronds dry roasted and cooked as part of hot dishes, this is known as "cochayuyo". (SANTELICES, 1989). The main production areas are in regions VII and X. (RED, 1990).
FUENTE: http://www.angelfire.com/sd/Lasalgas/itcl001.html
Algae most important in Chile
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PRODUCT
VOL 2010 (Ton)
Value 2011 (USD Mill)
Algas (total) 65100 81,2
Lessonia (chascon) 52800
Macrocristis (Huiro) 3300
Gracilaria (pelillo) 2900
Carrageninas 5210 52,9
Agar Agar 2170 43,7
Alginatos 1756 20,6 Fuente: Intern. Trade Center – Trade Map
Description of exportable supply of Chile
8
Chilean production of different algae and algal products for export FOB amount reached close to USD 200 million, with volumes close to the 75,000 tons.
Export supply of Chile, the most important product in volume are natural dried seaweed, especially the genus Lessonia. However, when analyzing the export earnings, are Derivatives from Algae who take the utmost importance and represent 59% of the returns.
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium Tabla: Exportaciones de algas pardas 2010. FUENTE: Revista Aqua. Mayo 2011.
Description of exportable supply of Chile
The kelp industry has had a high growth worldwide and in this area, Chile contributes with 10% of the biomass. Of these, the ones with the greatest commercial importance are Lessonia nigrescens, Lessonia trabeculata, Macrocystis integrifolia and Macrocystis pyrifera Durvillacea Antarctic.
Some of these species are of great economic importance, since they are extracted aliginatos used in the pharmaceutical industry, food (animal and human), textile, biomedical and cultivation of land plants, among other things.
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
China66%
Japan10%
France9%
Norway7%
Denmark3%
Others5%
Target Markets - ALGAE
Target markets: natural seaweed
10
The main destination markets for natural seaweed production in Chile in 2011 were China (with 67% of the tonnage shipped), Japan, France and Norway.
Different markets have virtually remained the volumes imported during the last 5 years, except China, which doubled its imports from Chile in this period.
Chile has had an average annual growth rate of 7% in these exports over the past 5 years.
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Seaweed products
PHYCOCOLLOIDS
Definition: Colloids are complex polysaccharides capable of forming gels, viscous substances and stabilizers of suspensions, according to its concentration and type of colloid (agar, vegetable gums, starch, Peptinas, alginates, carrageenans, etc.). The FICO prefix means Algae (Latin).
Alginates or alginic acids: Alginic acid obtained from different types of algae (Macrocrystis, Lessonia, Fucus, etc.). Extracting with sodium carbonate and precipitating with acid treatment. The gels are formed in the presence of calcium, which must be added in a controlled manner to achieve the formation of ordered molecular associations are not reversible by heating, this property makes alginates unique among all gelling agents. It is used in canned vegetables and jams, confectionery, cakes and cookies and ice cream. Also used the development of cold meats, pates, dehydrated soups, to keep the pulp suspension in fruit nectars and soft drinks that contain as foam stabilizer in beer.
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Seaweed products
12
Carrageenan: Are obtained from various types of red algae (Gigartina, Gracilaria, Furcellaria). The carrageenans are acidic and form thermally reversible gels, and it is necessary to dissolve in hot. They are widely used in the preparation of desserts, and they interact very favorably with milk proteins. With concentrations of 0.025% of carrageenan, it can stabilized suspension and at around 0.15% already provide solid textures .
Agar-agar: In Chile It is obtained mainly from Gracilaria chilensis. Is a polysaccharide widely used in microbiology, as an excellent growth medium for microorganisms.
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Olmix Algae Symposium - Sept. 10th, 2012
Japan34%
USA20%
Russian Fed18%
Thailand7%
Others21%
Target markets - AGAR AGAR
Target markets: products of algae
13
Even when China is the largest importer of natural algae in Chile, that country is not important destination of the processed products derived from algae.
In the case of agar, and alginates, the main countries receiving exports from Chile are Japan and USA.
In the case of carrageenan, USA and Denmark are the main destinations of exports from Chile.
France is an importer of Natural Seaweed, alginates and carrageenan with a 9%, 6% and 8% respectively from the total export volumes from Chile.
Japan26%
USA20%
Brazil16%
Mexico12%
France6%
Others20%
Markets - ALGINATES
USA23%
Denmark22%
Mexico15%Spain
6%France
3%
Others31%
Markets - CARRAGEENAN
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Producto
Precio prom (USD/Kg)
Rango según importador
Agar Agar (2010) 16,8 14,7 - 18,1
Carragenina (2011) 10,0 7,9 - 11
Alginatos (2010) 14,2 11,3 - 16,2
500
700
900
1 100
1 300
1 500
1 700
1 900
2007 2008 2009 2010 2011
Prices: ALGA NATURAL (USD/Ton)
China
Japan
France
Promedio
Export Price
14
In 2011, the average export price of algae in the wild was 1.193/Ton USD (FOB).
This price has shown an average annual increase of about 11% over the past 5 years, however the trend over the past 2 years has been to stabilize.
Among processed products derived from seaweed, Agar Agar shows a higher average price per kg (USD 16.8), followed by Alginates and Carrageenan.
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Conclusions from the point of view of Chile's export supply
1. There has been a sharp increase in world demand for Chilean supply of algae in their natural state and also for products derived from algae, due to its quality and export capacity in the country.
2. Chile has shown a very quick and efficient ability to increase their export volumes to meet rising world demand, yet there is great potential for further growth in the country.
3. The export supply in Chile is not only crude feedstock (algae naturally), but also products derived from algae, especially Agar Agar, Alginates and Carrageenan.
4. The export prices of Chilean Offer are convenient, especially for tariff benefits with importers, thanks to the many trade agreements Chile has signed with most countries.
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EMPRESA DIRECCION TELÉFONO WEB
Prodalmar Ltda. Calle General Velásquez 890 Of. 602, Antofagasta .
(55) 284148; (55) 224949
http://www.prodalmar.cl/
ALIMEX S.A. Isidora Goyenechea 3621 · Piso 17 · Las Condes, Santiago.
(56 2) 4301200 http://www.alimex.cl/espanol/
Guangjin Ltda. Doctor Johow 672, Santiago. (56-2) 2765935 http://www.bentalina.cl
Exportaciones M2 s.a. Melgarejo 750 Of. 74, Coquimbo. (56) - (51) 341758
Algas Cruz Alta S.A. Félix de Amesti 124, Of. 41, Santiago. (56-2) 2455757 www.algascruzalta.com
Costa Azul S.A. (32) 2972593 http://www.ipesca.co.cl/
Cultivos Acex S.A. Mardoqueo Fernández 156, Providencia, Santiago.
(2) 2331304
Algas Vallenar S.A. Vallenar: Panamericana Norte S/N Km. 665
MJ Chile Ltda. Vía 1 Sitio 2 Km.10 - Bajo Molle, Iquique. 412044
Pampamar S.A. Av. Las Condes 8060 Of. 203, Santiago. (56-2) 2297223 http://www.pampamar.cl/
Inversiones Kelp S.A. GENERAL VELASQUEZ 890, OF. 803, Santiago
(56-2) 3353309
CULTIVOS MARINOS CHILESUR S.A
Pacheco Altamirano, pasaje Canal Tenglo N° 2932, Puerto Montt
Terra Natur S.A. Hualqui (41) 2780987
Algae companies in Chile
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Chilean regulations
CHILEAN FOOD STANDARDS Chile has regularized within the aquaculture products, in the Chilean Norm 1857. Office 84, this standard applies to the algae commonly known in Chile for commercial purpose, with common and scientific nomenclature.
FOOD HEALTH REGULATIONS According to Health Regulations of food, derived from seaweed (phycocolloids) are considered as food additives as required by Title III, Paragraph I, Article 130, since they only meet a technological and nutritious. In Title III, Paragraph II The use of additives, Article 149 states that are allowed to use as thickeners and hydrocolloids with good manufacturing practices agar, alginate, ammonium, calcium and sodium, Carrageenan. In the provisions of Title II, Paragraph II of the labeling and advertising, Article 107, all food products must bear a label or tag on their packaging contain the following information: name of the product, net contents, name or business name and address of manufacturer, country of origin, number and date of the health decision, dates, and duration of the product, instructions for use and storage. (Food Health Regulations 1998).
17
Thanks for your attention!
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« Algae, source of active principles in cosmetics »
Alexis Rannou - Deputy Managing Director in charge of Innovation - ARD Soliance
1
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
La cosmétique Définition
[C’est une substance ou une préparation destinée à être mise
en contact avec les diverses parties superficielles du corps humain, notamment l'épiderme, les systèmes pileux et capillaire, les ongles, les lèvres et les organes génitaux externes, ou avec les dents et les muqueuses buccales, en vue, exclusivement ou
principalement, de les nettoyer, de les parfumer, d'en modifier l'aspect, de les protéger, de les maintenir en bon état ou de
corriger les odeurs corporelles.]
Source: article L.5131-1 du code de la santé publique 2
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
La cosmétique Composition d’un produit cosmétique
EAU 65-85%
Huiles 5-25%
• Ingredients & Actives 0,5-5% • Conservateurs 0,1-0,5% • Parfums 0,1-0,5%
3
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La cosmétique Structure de la peau
La peau: un organe de 2,5 m² constituée de 70% eau
Collagène + GAG
HA + Chondroitine sulfate
Epiderme: 13% eau
Derme: 57% eau
4
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Pourquoi les algues ? Une biodiversité exceptionnelle
1. Les algues, un potentiel d’innovation
Microalgues/phytoplanctons : Des milliers d’espèces dans plusieurs classes taxonomiques, très diversifiés du point de vue phylogénétique :
• Procaryotes = Cyanobactéries (autotrophes) • Eucaryotes = Algues vertes, rouges, dorées et brunes
Macroalgues : Pluricellulaires, attachés à des substrats :
• Vertes • Rouges • Brunes
5
NB : Les plantes supérieures comprennent plus de 250 000 espèces dont la plupart sont comprises dans une seule classe
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium 6
Pourquoi les algues ? Une biodiversité exceptionnelle
2. Une biodiversité de morphologies
Microalgues : • Seules ou en colonies (sous forment de chaine ou de filament) • Certaines sont mobiles grâce à leurs flagelles
Macroalgues :
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium 7
Pourquoi les algues ? Une biodiversité exceptionnelle
3. Une diversité des habitats et des environnements
Habitat : • Eau de mer • Eau saumâtre ou douce • Ou même les sols, les rochers et les arbres
Environnements : • Sources hydrothermales • Arctique et Antarctique • Zone aride • Forte pression osmotique • UV
Olmix Algae Symposium - Sept. 10th, 2012
Solution clarifiée (Sucres et
polysaccharides…)
Extraits peptidiques
Autres extraits (sels, pigments, oligoélements)
Extraits lipidiques
Pourquoi les algues ? Une biodiversité exceptionnelle
8
4. Grande disponibilité
• Sécurité d’approvisionnement (production: 14,7 Mn Tonnes de macroalgues en 2009)
5. Potentiel d’activités et de fonctionnalités
• Production de métabolites + fonction des conditions du milieu
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Solution clarifiée (Sucres et
polysaccharides…)
Extraits peptidiques
Autres extraits (pigments, sel, oligoélements)
Extraits lipidiques
Polysaccharides sulfatés Rhéologie HPM Activité biologique
MPM et BPM
Actions nutritives Actions peptides Amincissants, etc.
Antioxydants Zinc, Manganèse, etc.
Potentiel important sur AGPUFAs
Pourquoi les algues ? Une biodiversité exceptionnelle
9
Solution clarifiée (Sucres et
polysaccharides…)
Extraits peptidiques
Autres extraits (sels, pigments, oligoélements)
Extraits lipidiques
Potentiel important AntioxydantsActions nutritivesPolysaccharides sulfatés
5. Potentiel d’activités et de fonctionnalités
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Pourquoi les algues ? Une source d’innovation à haute valeur ajoutée
10
BIODIVERSITE ALGUES = ENORME POTENTIEL
Nombre d’espèces
Nombre de molécules
Conditions de culture
HAUTE VALEUR AJOUTEE
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Un besoin marché Les algues comme matières premières cosmétiques
Macroalgues
Ingrédient clé de la réussite de la cosmétique marine française, unique et reconnue dans le monde entier (instituts, thalasso, spas, masques, etc.)
Excellente dermo-compatibilité des algues
Rôle de texturant
Source de tensio-actifs naturels.
Source d’ingrédients actifs • Concernant les actifs cosmétiques issus de phycocolloïdes, on trouve les oligo-alginates
présentant des actions anti-pollution, anti-éruption cutanée, anti-acné, et antivieillissement.
• Les fucoïdanes agissent sur la réduction du vieillissement de la peau et favorisent la croissance capillaire.
11
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium 12
Un besoin marché Les algues comme matières premières cosmétiques
Microalgues
Cosmétique : un concept biomimétique
Anti-âge : puissants antioxydants (caroténoïdes, groupe d'enzyme SOD, les tocophérols ou l'ascorbate)
Protection UV : Synthèse des pigments anti-UV mycosporine et scytonemine (cyanobactéries «de l’extrême» du genre Nostoc*)
Hydratation : Synthèse d’exopolysaccharides en cas de sécheresse ou en présence d’un environnement agressif
Les microalgues sont riches en PUFAs : ils représentent une grande ressource pour des applications cosmétiques
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- Sélection après études bibliographiques
- Différentes provenances : algothèques privées, partenariats,
- Isolement sur site et identification de l’espèce (par PCR)
- Différents milieux : marines, eaux douces et eaux saumâtres
Centre de Biotechnologie Marine Des labos de R&D
Souches de microalgues
13
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
- Adaptation des souches au laboratoire
- Optimisation des paramètres de culture
- Cultures des microalgues jusqu’au stade du photobioréacteur de 300L
Centre de Biotechnologie Marine Un savoir-faire
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Photobioréacteurs de 2100L
2 Bassins de 55 m3
Récolte par ultracentrifugation
+ Partenariats avec des sites de production industriels (capacités > 1 T MS)
Centre de Biotechnologie Marine Une unité de validation industrielle et de production
15
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Quelques exemples Les algues comme matières premières cosmétiques
16
Quelques exemples d’actifs d’origine marine
Olmix Algae Symposium - Sept. 10th, 2012
Soliance : Wakamine
17
Wakamine
Eclaircissez votre peau
Puissant actif blanchissant d’origine marine
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
La pigmentation de la peau résulte de la présence de mélanine dans l’épiderme.
Wakamine Contexte biologique
18
Deux types de mélanine : • l’eumélanine, brun-noir, • et la pheomélanine, jaune-orange.
La tyrosinase est l’enzyme indispensable pour les deux premières étapes de la mélanogénèse. Wakamine est un actif blanchissant performant Il inhibe la tyrosinase, l’enzyme clé impliquée dans la production de mélanine.
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Plusieurs noms lui sont donnés : Chine : Qun dai cai (Lembi & Waaland 1988)
Angleterre : Sea mustard (Kirby 1953), Precious sea grass (Rhoads & Zunic 1978)
Japon : Wakame (Madlener 1977)
Corée : Miyok (Madlener 1977), Miyeouk (Lembi & Waaland 1988)
Source : Algaebase.org
Alanine 7.6%
Arginine 5.7%
Acide aspartique 9.5%
Cystine 2.3%
Acide glutamique 12.4%
Histidine 5.9%
Isoleucine* 4.3%
Leucine* 8.0%
Lysine * 3.5%
Methionine* 3.4%
Proline 5.9%
Serine 4.7%
Tryptophane* 1.5%
Threonine* 5.0%
Tyrosine 3.7%
*Acides aminés essentiels
Wakamine Undaria pinnatifida
19
Wakamine est extrait d’une macroalgue brune, Undaria pinnatifida, de l’ordre des Laminariales.
Cette algue présente un profil unique en acides aminés, avec plus de 42% d’acides aminés essentiels.
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Cette algue fut introduite en France en 1971 en Méditerranée (étang de Thau) et par IFREMER sur les côtes Bretonnes en 1983.
Wakamine Origine géographique
20
Undaria pinnatifida : Originaire des baies de la mer du Japon à l’ouest d’Hokkaido ainsi que des baies Coréennes et Chinoises, elle est traditionnellement cultivée et largement utilisée dans la cuisine asiatique.
Olmix Algae Symposium - Sept. 10th, 2012
Wakamine Culture de l’algue
21
Undaria pinnatifida est récoltée dans ses habitats naturels depuis des siècles, elle est également cultivée. Sa production est estimée entre 450 000 et 500 000 tonnes au Japon et en Corée.
Culture de l’algue sur l’île d’Ouessant Cette macroalgue est aujourd’hui cultivée sur l’île d’Ouessant en Bretagne, un site classé par l’UNESCO.
Olmix Algae Symposium
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En inhibant la tyrosinase, Wakamine limite la production de mélanine
Wakamine Mécanisme d’action
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Wakamine inhibe la production de mélanine : -39%
Témoin: les mélanocytes sont visibles
Wakamine (100mg/ml) : Les mélanocytes sont légèrement
visibles
Wakamine Inhibition de la production de mélanine (test ex vivo)
23
Protocole Les épidermes sont obtenus à partir d’explants de peau. Ils sont séparés après incubation dans du NaBr 2N pendant 100 minutes à 37°C. Les épidermes sont ensuite fixés avec une solution tampon de formaldehyde, rincés et traités avec un mélange de l’actif (100μg/ml) et d’une solution de DOPA (1mg/ ml) pendant 4 heures à 37°C. Après incubation, les épidermes sont rincés et visualisés au microscope. Une analyse quantitative est réalisée par traitement d’images.
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Wakamine aussi efficace que l’Arbutine (test in vivo)
24
Protocole Une solution de Wakamine 3% est appliquée sur les explants de peau à l’aide de patchs de papier Whatmann une fois par jour pendant 6 jours. Les explants sont fixés dans le formol et montés dans la paraffine. La coloration due à la mélanine sur les coupes est traitée selon la technique de Masson. L’intensité de couleur est calculée par analyse d’images.
Wakamine est plus performant que l’Arbutine (référence quasi drug au Japon)
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Wakamine éclaircit la peau (test ex vivo)
25
-10
-8
-6
-4
-2
0J0 J28 J56
Varia
tion
de
l'Ind
ex m
élan
ique
(eff
et e
clairc
issan
t)2% Arbutine
1% Arbutine + 1% Wakamine
* p< 0.001 Chaque valeur de l’index mélanique est significativement différente par rapport à la valeur de J0.
* *
La diminution est effective pour 91% du panel
Protocole L’efficacité éclaircissante de Wakamine a été évaluée sur un panel de volontaires de type asiatique (22 volontaires, d’âge moyen 43 ans). Les panélistes appliquent 2 fois/jour pendant 56 jours.
En remplaçant 1% d’arbutine par 1% de Wakamine, des résultats in vivo similaires sont obtenus.
Wakamine permet de rendre vos formules plus innovantes
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Soliance Microalgue - Phaeodactylum tricornutum
MEGASSANE Protection ciblée des protéines contre les rides
26
Extrait d’une microalgue appartenant à la famille des Diatomées.
Microalgue unicellulaire très répandue dans les eaux côtières et saumâtres des zones tempérées
Actif breveté (prix nobel) - Anti-photovieillissement
• Maintien de l’homéostasie de la peau • Purification des cellules • Nouvelle approche pour lutter contre le vieillissement.
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Soliance Microalgue - Phaeodactylum tricornutum
Mécanisme d’action : Détoxifications des cellules Mégassane stimule les trois activités du
protéasome et les restaure après exposition aux UV pour limiter l’oxydation des protéines
Megassane est recommandé dans : Des produits anti-âge Protections solaires Gammes de soins purifiants/détoxifiants
27
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Soliance Microalgue - Phaeodactylum tricornutum
28
Stimulation des activités du protéasome (test in vitro) Protocole Les kératinocytes sont traités avec 2,5 μg/ml d’extrait de P.tricornutum pendant 24 h. Les activités peptidases du protéasome sont évaluées dans le lysat cellulaire en utilisant 25 μM de LLVY-amc, 150 μM de LLE-na, et 40 μM de LSTR-amc comme substrat peptidique fluorescent avec 20 μg de protéines totales.
En stimulant les activités du protéasome, Megassane diminue le taux de protéines oxydées
Diminution du taux de protéines oxydées (test in vitro) Protocole Un Oxyblot est réalisé avec 10 μg de protéines issues du lysat de kératinocytes, cultivés 7 h après traitement avec 2,5 μg/ml d’extrait de Phaeodactylum.
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Soliance Microalgue - Phaeodactylum tricornutum
29
Restauration des activités du protéasome après exposition aux UV Protocole Une culture de kératinocytes est irradiée par des UVA UVB et/ou traitée par de P.tricornutum pendant 24 h. Les activités peptidases du protéasome sont évaluées dans le lysat cellulaire.
En restaurant les activités du protéasome Megassane répare les protéines endommagées par les UV
Réparation des dommages protéiques dus aux UV Protocole Une culture de kératinocytes, préalablement exposée aux UV, est traitée avec 2,5 μg/ml d’extrait de Phaeodactylum. Un Oxyblot est réalisé avec les protéines issues du lysat.
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Complexes Argilo-Algales Brevet Mondial Amadéite®
30
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Revertime®
Antirides et anti-âge exceptionnel
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Revertime® Antirides et anti-âge exceptionnel
Forte activité anti-oxydante protégeant efficacement la peau des attaques radicalaires occasionnées par le stress de la vie courante :
32
- 52% * de réduction de réaction anti radicalaire (* à 0,25%)
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Revertime® Antirides et anti-âge exceptionnel
33
Activité anti-élastastique hors du commun : pouvoir inhibiteur de l’élastase (contribue à la dégradation des fibres élastiques):
- 100 %* de réduction de la dégradation des fibres élastiques (* à 0,1%)
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Revertime® Antirides et anti-âge exceptionnel
34
- 64% * synthèse collagène type 1
(* à 0,01%)
Forte production de pro collagène de type 1 l’un des constituants majeurs du derme :
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Revertime® Antirides et anti-âge exceptionnel
35
La surface et le volume des rides : -12,3 % L’amplitude et la rugosité moyenne du tissu cutané : -5,5 et -2,6 %
In vivo, effet sur les rides de la pate d’oie après seulement 28 jours, réduction significative de :
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Revertime® Antirides et anti-âge exceptionnel
Un actif antirides et anti-âge exceptionnel. Très facilement incorporable en formulation, Revertime® propose une solution efficace et pertinente dans la lutte contre le vieillissement cutané.
36
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Emultime®
Auto Emulsifiant naturel, technologie à froid
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Emultime® Auto Emulsifiant naturel, technologie à froid
Sous simple agitation à froid ou à chaud, Emultime® permet d’obtenir des mélanges « Eau/huile » homogènes et stables
Gain énergétique considérable par rapport à une émulsion classique à chaud
Mise en œuvre plus rapide et process simplifié : temps de production divisé par 4
100% naturel : approche écologique de la formulation cosmétique
Permet de réaliser des laits et des crèmes cosmétiques avec des textures originales
38
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Conclusion
39
Les algues sont les MP cosmétique
du présent et du futur car elles ont
un véritable potentiel de découverte
en terme d’origine, d’activité et de
fonctionnalité.
88
« Algae, source of nutriments for humans »
Dr Maria Hayes - Researcher and Principle Investigator of NutraMara Biodiscovery and characterisation and Scientific Programme Manager (2008-2012) -
Teagasc - The Irish Agricultural and Food Development Authority
1
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Teagasc Food Research Centres
Teagasc Nutraceutical Research Facility
Introduction
2
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State of the Art Teagasc represents in Ireland
3
“Provide a resource base of knowledge and skills to stimulate the innovation function”
Teagasc Ashtown Food Research Centre Teagasc Moorepark Food Research Centre
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Challenges facing stakeholders
4
Legal restrictions, high costs and environmental problems regarding the disposal of marine processing wastes.
European legislation (EU Council Directive 1999/31/EC, 1999) specific targets for disposal at landfills.
The Marine Functional Foods Research Initiative focuses on three main marine resources (i) fish processing waste streams, (ii) seaweeds and microalgae and (iii) aquaculture sources.
The Marine Functional Foods Research Initiative aims to exploit these marine resources for functional foods/ ingredients development.
Identification of bioactive compounds to the dietary intervention level in order to make a health claim.
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Very diverse pool of potential food ingredients from aquatic sources
5
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Significant interest in marine proteins
6
Fish protein Gelation
Water binding
Solubility
Emulsification Foaming
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Market Trends The global market for functional foods $174 billion.
Functional foods and nutraceuticals offer opportunities for the agri-food sector… to become the ‘pharmacy for disease prevention’.
7
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Key Areas based on Market and Scientific Opportunity
8 Olmix Algae Symposium
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Traditional use of Seaweed in Ireland and worldwide
During Potato famine seaweed source of nourishment.
Sluichèan: a seagrass.
High nutrient content.
Dulse Bread: red seaweed.
Seaweed : in Asian countries for centuries in foods, such as sushi.
9
Dulse Bread
Sluichèan
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Traditional use of Seaweed
“Nori” - Porphyra sp. (Sushi)
“Habi-Nori” - Petalonia binghamiae (Eaten dried and roasted).
“Hijiki” - Hizikia fusiforme (Soups, salads, vegatable dishes).
“Kombu” - Laminaria sp. (dashi soup)
“Wakame” - Undaria pinnatifida (Salads, soups).
10
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Harvesting To the Dish
Animal products
Traditional use of Seaweed
11
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(FAO, 2010)
Macroalgae production world-wide
12
Harvested - 1,045,000 T ww Cultivated - 15,781,159 T ww
CHINA, top seaweed producer • CHINA 62.8% • INDONESIA 13.7% • PHILIPPINES 10.6% • REPUBLIC OF KOREA 2.8% • JAPAN 2.9%
JAPAN is the second most important producer in terms of value = NORI production.
CHILE 21,700 T
Saccharina japonica, CHINA
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Open-water system in Canada (IMTA)
Macroalgae production world-wide
13
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
New derivatives
Development of
extractions
Defining activities
Marketing and product development
Proteins, peptides and amino acids
Seaweed sources H H H H
Seafood sources H H H H
Carbohydrates
Chitin and chitosan L M H H
Agar L L L M
Carrageenan L L M M
Alginate L L L M
Fucoidan H H H H
Laminaran M M M M
Ulvan M M M M
Antioxidants M H H H
Omega-3 fatty acids M H M H
Innovation potential summary
14
H = high potential
Olmix Algae Symposium - Sept. 10th, 2012
Algal bioactive ingredients
15 HO
OH
HO
O
HO
OHOHO
O
HO
HO
HO
OHHO
n
Antioxidants
Blood pressure Cancer Inflammation Obesity Diabetes Mental health
Laminarin
Fucoidan
LIPIDS
93
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Bioassays : Renin Inhibition
Aspartyl protease.
Sodium depletion, decreased blood volume and blood pressure and β-adrenergic stimulation.
Angiotensin II inhibits renin secretion and has a number of physiological effects.
Cleavage of angiotensinogen by renin is the rate determining step in the RAS.
Inhibition of renin attractive strategy for control of hypertension.
16
RAAS system
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Platelet Activating Factor – Acetylhydrolase (PAF-AH) Inhibitors
Plasma PAF-AHs are located cytosolic and plasma.
Selective for phospholipids with very short acyl groups.
Linked to atherosclerosis and may be a positive risk factor for coronary heart disease in humans.
Screening for inhibitors of this from seaweed and marine extracts and bovine muscle extracts.
Process of atherosclerosis
Platelet-activating factor acetyl-hydrolase 17
Olmix Algae Symposium - Sept. 10th, 2012 18
Bioactive peptides sources
Marine by-products and algal sources…
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae
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Generation of bioactive peptides
Characterisation – UPLC & MS
Freeze-dry
Freeze-dry
Extraction of macroalgal, microalgal or marine by-product proteins
Hydrolysis with proteolytic enzymes
Purification RP-HPLC
Test hydrolysates for PAF-AH, PEP, ACE-I and renin inhibition
UF 10kDa MWCO/3kDa MWCO
19
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Background
Hypertension is one of the major yet controllable risk factors in CVD but it can be controlled by inhibition of a number of enzymes in the Renin Angiotensin System (RAS).
20
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Background
Platelet activating factor acetylhydrolase (PAF-AH) is a circulating enzyme produced and secreted by inflammatory cells centrally involved in atherosclerosis.
It generates two key pro-inflammatory mediators, lysophosphatidylcholine (LPC) and oxidized nonesterified fatty acids (oxNEFAs).
Evidence exists for a regulatory role of these lipids in promoting atherosclerotic plaque development that can ultimately lead to the formation of a necrotic core, a key determinant in atherosclerotic plaque vulnerability.
21
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Algal proteins and peptides inhibit Renin
Isolation of protein from Palmaria palmata
Generation of peptides using food grade enzymes
Enrichment of peptide fractions and purification of peptides
Characterisation of bioactive peptides (bioassay and UPLC-MS)
22
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Protein extraction
Crude protein was extracted from Palmaria palmata using the method previously described by Galland-Irmouli.
Following a cold water extraction the protein fraction of was precipitated with ammonium sulphate.
23
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Generation and enrichment
24
0.00
10.00
20.00
30.00
40.00
50.00
60.00
70.00
Fr10 Fr11 Fr12 Fr13 Fr14 Fr15 Fr16 Fr17 Fr18 Fr19 Fr20 Fr21 Fr22 Fr23 Fr24 Fr25
RP-HPLC fractions
Perc
enta
ge In
hibi
tion
RP-HPLC
Renin bioassay 58.97 % inhibition (+/- 1.26) IC50 0.32mg/ml
F10-F25
96
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Sequence Phylum Protein (assession number) Species sharing 100% homology
Calculated mass
Observed mass
Charged state
D.IRLIIVLMPILMA.A Rhodophyta Photosystem II protein Y (O19893)
Cyanidium caldarium,Galdieria sulphuraria 1494.93 499.29 (+3)
MNEIVALMI.I Rhodophyta Cytochrome b6-f complex (Q85FX8) Cyanidioschyzon merolae 1032.53 517.26 (+2)
P.ILMA.A Rhodophyta Photosystem II protein Y (O19893)
Cyanidium caldarium,Galdieria sulphuraria 446.25 447.1235 (+1)
I.LMAASWAIY.N Rhodophyta Photosystem II protein Y (O19893)
Cyanidium caldarium,Galdieria sulphuraria 1024.5 1025.48 (+1)
Q.ILPSILVPLV.G Rhodophyta Photosystem I reaction center subunit VII (P58214) Chlorella vulgaris 1062.7 532.32 (+2)
L.PSIL.V Rhodophyta Photosystem I reaction center subunit VII (P58214) Chlorella vulgaris 428.26 429.09 (+1)
I.LVPLVGLV.F Rhodophyta Photosystem I reaction center subunit VII (P58214) Chlorella vulgaris 808.54 809.4 (+1)
V.PLVGLVFPAI.A Rhodophyta Photosystem I reaction center subunit VII (P58214) Chlorella vulgaris 1024.63 1025.48 (+1)
L.VFPAIAM.A Rhodophyta Photosystem I reaction center subunit VII (P58214) Chlorella vulgaris 747.39 748.4 (+1)
V.FPAI.A Rhodophyta Photosystem I reaction center subunit VII (P58214) Chlorella vulgaris 446.25 447.12 (+1)
F.PAIA.M Rhodophyta Photosystem I reaction center subunit VII (P58214) Chlorella vulgaris 370.22 371.11 (+1)
Peptide identification by tandem mass spectrometry
25
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Renin inhibition assay of synthesised peptides
Sequence Renin inihibition +/-
D.IRLIIVLMPILMA.A 51.38 3.5
MNEIVALMI.I - -
P.ILMA.A 29.34 4.59
I.LMAASWAIY.N - -
Q.ILPSILVPLV.G 24.56 3.56
L.PSIL.V - -
I.LVPLVGLV.F 15.35 2.16
V.PLVGLVFPAI.A 17.5 3.21
L.VFPAIAM.A - -
V.FPAI.A - -
F.PAIA.M - -
26
IC50 (mM)
3.34
Fitzgerald & Hayes, 2012, J. Agriculture and Food Chemistry
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Bioassays: PAF-AH inhibition
PAF-AH is an enzyme that converts PAF to the biologically inactive lyso-PAF.
PAF is a biologically active phospholipid that is involved in activation of platelets, monocytes, macrophages, polymorphonuclear leukocytes.
PAF-AH is linked to atherosclerosis and may be a positive risk factor for coronary heart disease in humans.
27
Figure 1: PAF-AH inhibitory activities presented as percentage inhibitory values for the trout myofibrillar protein thermolysin 10-kDa-UFH RP-HPLC fractions.
PAF-AH inhibition by RP-HPLC fractions generated from trout myofibrilar muscle hydrolysed with thermolysin and filtered
through a 10-kDa membrane filter
020406080
100120140
MAFP control F22 F10 F26 F25
Sample name
% P
AF-
AH
inhi
bitio
n
97
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Bioassays: PEP inhibition
0102030405060708090
100
Berberine Trout fullhydrolysate
3kDa trout fitrate 10kDa troutfiltrate
Inhibitory sample name
% In
hibi
tion
of B
SA
28
Enzyme prolyl endopeptidase (PEP) plays a role in the degradation and metabolism of biologically active peptides containing proline such as oxytocin, vasopressin, substance P, bradykinin, neurotensis, and angiotensins .
Important biological functions in organs such as the brain and have been implicated to play a role in the development of neurodegenerative conditions such as AD.
Furthermore, specific inhibitors of PEP have anti-amnesic effects, and some of them have been synthesized as anti-amnesic drugs.
Figure 2: PEP inhibitory activities presented as percentage inhibition values for the trout myofibrillar protein thermolysin hydrolysates, 10-kDa and 3-kDa filtrates.
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Mass spectrometry analysis and determination of peptide amino acid sequences
0
20
40
60
80
100
%Int.
800 1000 1200 1400 1600 1800 2000 2200 2400 2600 2800 3000Mass/Charge
1[c].M22
45 mV[sum= 7655 mV] Profiles 1-171 Unsmoothed
Data: Actin 2 Hours0001.K19 16 Jan 2008 20:36 Cal: 160108_2 16 Jan 2008 20:23 Kratos PC Axima CFRplus V2.4.1: Mode reflectron, Pow er: 79, Blanked, P.Ext. @ 3000 (bin 132)
1827.83 {r4723}
2482.81 {r7464}
2394.94 {r8578}
720.08 {r6353}2406.82 {r4960}
1433.66 {r3009}1021.47 {r4537}2221.90 {r7462}1458.54 {r6203} 2504.81 {r5608}716.56 {r11121} 1093.47 {r3743} 1823.74 {r7250} 2866.00 {r9638}2218.80 {r10980}
29
Fingerprint Q-TOF MS and UPLC system
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Peptide characterisation
Protein identified using Protein Lynx Global Server 2.4
and TurboSEQUEST
Peptide sequence Position Peptide mass (Da)
Charge state
Protein Lynx Global Server 2.4 & peptide
source
TurboSEQUEST and peptide source
Hemoglobin subunit beta
FGKEFTPVLQADFQK 117-131 1754.911 (3+) 10-kDa_UFH Full hydrolysate
Hemoglobin subunit beta
FGDLSTADAVMNNPK 44-58 1579.742 (2+) 10-kDa-UFH RP-HPLC fraction 3 RP-HPLC fraction 4 RP-HPLC fraction 6
Hemoglobin subunit beta
LHVDPENFKL 95-104 1211.641 (2+) 10-kDa-UFH RP-HPLC fraction 3
Hemoglobin subunit alpha
VLSAADKGNVKA 2-13 1172.663 (2+) 10-kDa-UFH Full hydrolysate
Catalase FSDVHPEYGSR 482-492 1293.585 (2+) 10-kDa-UFH Full hydrolysate
Catalase AQKPDVLTTGGGNPVGDKLNS 21-41 2068.066 (2+) 10-kDa-UFH Full hydrolysate, RP-HPLC fraction 3
Catalase AAQKPDVLTTGGGNPVGDKLNS 20-41 2139.104 (2+) 10-kDa-UFH Full hydrolysate
Catalase LVQDVVFTDEMAH 51-63 1503.714 (2+) 10-kDa-UFH Full hydrolysate
Fatty acid-binding protein
VGMPDDIIQKGKD 22-34 1415.719 (2+) 10-kDa-UFH Full hydrolysate
30
**Mora & Hayes, 2012, Identification of Peptide Sequences and Bioactive Characterization of Rainbow Trout Thermolysin Hydrolyzate Fractions with in vitro Inhibitory activities against Enzymes Important in Heart and Brain Health (Accepted in the Journal of Agriculture and Food Chem)
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Olmix Algae Symposium - Sept. 10th, 2012
EPA (20:5 n-3)
Linolenic acid (18:3 n-3)
Palmitic acid (16:0) Palmitoleic acid (16:1 n-7)
Fatty acids Reduce risk of cardiovascular diseases
Possible health effects
Oleic acid (18:1 n-9) Antioxidant activity
Antioxidant activity
Antimicrobial activity Reduce risk of cardiovascular diseases
DHA (22:6 n-3) Improve childhood cognitive development
Reduce risk of old-age diseases such as Alzheimer Cancer prevention
Antiviral, antitumoral, antihyperlipidemia and anticoagulant
Sulfated polysaccharides
Insoluble fibers
Polysaccharides Possible health effects
Reduce total and LDL cholesterol
Potential functional products
31
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Generate accelerated solvent extracts (ASE®) from algal species
32
(5 min.)(5 min.)
Flush with fresh solventFlush with fresh solvent(0.5 min.)(0.5 min.)
Extract readyExtract ready(total time, 12(total time, 12
Load cellLoad cell
Fill with Fill with solvent(ssolvent(s))(0.5(0.5--1 min.)1 min.)
Static extractionStatic extraction(5 min.)(5 min.)
Flush with fresh solventFlush with fresh solventFlush with fresh solventFlush with fresh solventFlush with fresh solvent(0.5 min.)(0.5 min.)(0.5 min.)(0.5 min.)
Flush with fresh solventFlush with fresh solvent(0.5 min.)(0.5 min.)
Extract readyExtract readyExtract readyExtract readyExtract ready(total time, 12(total time, 12(total time, 12(total time, 12
Extract readyExtract ready(total time, 12(total time, 12--14 min14 min.)
Heat and pressuriseHeat and pressurise(5 min.)(5 min.)
Purge with nitrogenPurge with nitrogen(1(1--2 min.)2 min.)
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Hydrophilic ASE® extracts generated using ethanol: water 70:30 (v/v)
Code Name Date Location Sample weight
ISCG0365 Ascophyllum nodosum 08/02/2012 Finavara Co. Clare 100g DW
ISCG0238 Fucus vesiculosus 16/06/2011 Finavara Co. Clare 100g DW
ISCG0259 Fucus serratus 18/07/2011 Spiddal, Co. Galway 100g DW
ISCG0239 Fucus spiralis 16/06/2011 Finavara Co. Clare 100g DW
ISCG0257 Laminaria digitata 18/07/2011 Spiddal, Co. Galway 100g DW
ISCG0356 Ulva Intestinalis 24/01/2012 Spiddal, Co. Galway 750g WW
ISCG0355 Pelvetia caniculata 24/01/2012 Spiddal, Co. Galway 750g WW
ISCG0223 Fucus vesiculosus 08/06/2011 Golf Course, Galway 34 Kg WW
ISCG0072 Fucus vesiculosus 16/04/2012 Spiddal, Co. Galway 220g DW
ISCG0070 Cystoseira nodicaulis 30/03/2010 Finavara Co. Clare 42 g DW
ISCG0029 Codium fragile 02/02/2009 Spiddal, Co. Galway 15.75 g DW
ISCG0071 Palmaria palmata 04/11/2010 Spiddal, Co. Galway 70 g DW
33
99
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Using NMR for metabolite screening
34 Test extracts for antioxidant activities
Olmix Algae Symposium - Sept. 10th, 2012
Using 2D-NMR for metabolite identification
35
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Lipophilic ASE® extracts generated using chloroform: methanol 2:1 (v/v)
Code Name Date Location Sample weight
ISCG0356 Ulva intestinalis 08/02/2012 Finavara Co. Clare 100g DW
ISCG0269 Pelvetia canaliculata 16/06/2011 Finavara Co. Clare 100g DW
ISCG00223 Fucus vesiculosus 18/07/2011 Spiddal, Co. Galway 100g DW
ISCG0324 Fucus spiralis 16/06/2011 Finavara Co. Clare 100g DW
ISCG0353 Ascophyllum nodosum 18/07/2011 Spiddal, Co. Galway 100g DW
ISCG00283 Cytoseira tamariscofolia 24/01/2012 Spiddal, Co. Galway 750g WW
ISCG0029 Codium fragile 02/02/2009 Spiddal, Co. Galway 15.75 g DW
ISCG0071 Palmaria palmata 04/11/2010 Spiddal, Co. Galway 70 g DW
36
100
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Lipophilic ASE® extracts generated using chloroform: methanol 2:1 (v/v)
Extraction conditions
5 min preheat time 1,500 psi 120ºC Heat time 5 minutes Flush volume, 50 % of cell volume Purge time 60 s Static cycle : 4
37
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
ASE® extracts have been enriched for phlorotannins
Fucus serratus ISCG0068 ASE® M16 (5C)
Ascophyllum nodosum ISCG0032 ASE® M15 (1B)
Fucus vesiculosus ISCG0023ASE® M17 (3B)
A nodosum ISCG0032 ASE® M17 (9B)
C. nodcaulis ASE® M17 (10B)
The figure in brackets refers to the phlorotannin enriched extract.
Phlorotannin enriched extracts were examined using Q-TOF-MS.
38
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
UPLC-Q-TOF-MS of ASE® enriched phlorotannin extracts
Heptamer of phloroglucinol identified but not pure (745.1).
Also contains Iodine (126.9).
Quinic acid derivative of Phloroglucinol (316).
Fingerprint of all samples obtained (10 in total).
Further purification required.
39
101
Olmix Algae Symposium - Sept. 10th, 2012
Impact of Research
40
Support small/medium sized
enterprises
They would provide a novel source of bioactives for inclusion into Irish diets.
Improve Irish publics health
May help lower the rate of high blood pressure.
Contribute new ingredients to the growing Irish and
International functional food industry
May encourage people to buy indigenous food products.
Ensure protection of the Irish coastline.
Health-promoting bioactives from
seaweed
Environmental benefits
Thanks for your attention!
41
102
« Algae, a “new” nutrient resource for human food »
Christine LE TENNIER - C.E.O - Globe Export - Rosporden
1
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Founded on pioneering spirit and internationally oriented, Globe Export has specialised in the sale and transformation of nutritional marine plants for more than 25 years.
Used for ages, these plants are to expand their role in human nutrition over the next 15 years, thanks to their natural nutritional properties.
We develop healthy, nutritional, and tasty products which exceed expectations in terms of taste and nutrition.
High in protein, trace-elements and omega 3, combined with a low fat content, seaweed is a food source which can’t be beaten.
We cultivate and promote innovation & originality, thereby driving our development.
Algae: A “new” nutrient resource for human food
Globe Export, a vision for a future based on nutritional marine plants
2
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Based in Brittany, in an unspoiled setting including one of the largest natural seaweed fields in the world, providing extremely high-quality raw materials, Globe Export covers all aspects of the food production process
Small-scale production / trade: Algues de Bretagne®
Research and development / innovation: Creativ'Concept®
Algoculture section department: Globe Sea Garden®
The key figures:
3 industrial sites (seaweed, pasta, molecular cuisine) 100 SKUs 20 employees 2 Millions € T.O. (2011)
Algae: A “new” nutrient resource for human food
3
Globe Export, a vision of a future based on nutritional marine plants
103
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Dulse Palmaria palmata
Haricots de mer Himanthalia elongata
Laitue de mer Ulva lactuca
Nori Porphyra ombilicades
Wakamé Undaria pinnatifida
Kombu breton Laminaria digitata
With Algues de Bretagne, you can go beyond traditional nutrition.
Algae: A “new” nutrient resource for human food
4
Algues de Bretagne is the specialist in edible seaweed, with a wide range of products providing a plethora of tastes and ensuring high nutritional value.
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Algae: A “new” nutrient resource for human food
With its three separate ranges, Algues de Bretagne brings its specialist’s expertise to the table.
1. Seaweed range : Raw material for niche products: Seaweed tartare, marine spices, glasswort and sea ”beans”, fish rillettes with seaweed, seaweed chutney, fresh salted seaweed, fish soup and seaweed toast
5
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Algae: A “new” nutrient resource for human food
6
2. Pasta range:
Seaweed pasta using original shapes and tastes:
Dulse, Nori, Spirulina Flours.
Raspberry, cranberry, lemon-bergamot
Spicy colours, red beetroot, curry and Tomato.
Salmon ravioli.
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Algae: A “new” nutrient resource for human food
7
3. Molecular range:
Flavour Pearls range (solid on the outside with a liquid centre which bursts in your mouth). Available in ambiant T° with a shelf life of one year or in frozen form(IQF) with a shelf life of 12 months at -18°C.
Flavour Clouds range (dried arometized oils melt in the mouth), and Emulsions range (vinaigrette, citrus bergamot, Tomato Coriander base).
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Algae: A “new” nutrient resource for human food
Marine products are the future of the food industry
Seaweed provides an inexhaustible source of minerals and
high-quality nutrients.
Taste the difference delivered by Globe Export’s research into
the nutritional qualities of marine plants.
Our products and concepts represent a progressive, humanist
approach, a commitment to innovation and to the development of
marine products, which can meet the challenges of both taste
and health, so as to be fit for human consumption.
8
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Globe Export Zi de Dioulan - BP 37 - 29140 Rosporden - France
Tel: +33 (0)2 98 66 90 84 - Fax: +33 (0)2 98 66 90 89 www.algues.fr
Algues de Bretagne: where the sea and gastronomy meet
Algae: A “new” nutrient resource for human food
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105
« Seaweeds for soil and plant nutrition »
A fertilizer producing company case study
PRP Technologies - Dr. Bruno DARIDON - R&D manager 1
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Position
PRP offer products for sustainable agriculture.
Design, produce and distribute products & services that improve soils vital functions and that stimulates plants, allowing an optimized use of natural resources.
2
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium 3
The challenge : world agriculture has to face an increasing demand.
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Agriculture is a main issue
4
1,8
2,8
6,3
9
0
1
2
3
4
5
6
7
8
9
10
1900 1965 2000 2050
Mill
iard
s in
habi
tant
s
Year
World population
Source: Population Division of the Department of Economic and Social Affairs of the United Nations Secretariat (2007)
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Forcast
5
Increase neads for world production + 70% in 2050 + 1 milliard tons cereals and + 200 millions tons meat
Source: FAO
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Forcast
10% « only » of the target can be acheived by surfaces increase.
90% of the target has to be achieved by Yield improvement & intensification.
Source: FAO 6
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What does this chalenge means?
7
… Need of efficient soils with good fertility.
… Need of healthy plants, with high producing potential.
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Constraints on the chalenge !
8
Produce with less :
Produce better :
Energy
Water
Fertilizers
Agrochemicals / pesticides
Preserve biodiversity Limit EGS Protect soils Preserve water ressource quality
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium 9
Seeweads interests and their input towards the
agriculture challenges
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Some R&D sustainable directions for seaweed uses in agriculture
10
Solid or liquid products for improving seed germination, root development together with a good rhizosphere microorganism-plant synergy.
Liquid products for improving photosynthesis and plant resistance to both biotic and abiotic stresses.
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Seaweed in agriculture: an old story
11
Ancestral raw-material used since a long time in the fields near the coast areas.
All seaweed types (red, brown, green) where and are used. No known risk (toxic or ecotoxic) based on historical references and
long time use. Natural : -> Cosmetic, Food, Feed, Agriculture. Pérennial ressource if turnover & harvest are well managed. Composition:
– Nutritional interest for plants – Stimulation of natural plant defenses systems
-> How to developp in agriculture?
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Several raw-materials for several market segments
12
Cost + Added value + biological activity Concentration
Seaweed compost – Partly stabilized but some lossess of C, N & S by lixiviation + gaz emissions (EGS) + losses
of biological activity.
Fermented seaweed « juce » – Difficult to manage + partial losses of biological activity
Whole seaweed, fresh or dried – Quality + preserved biological activity
Seaweed refined fractions obtained by downstream processing technologies
– Liquid extract -> polysaccharides, proteins, native soluble metabolites – Hydrolysates -> oligomers with controlled DP & DS (biological activity). – Purified fractions: UF, NF, OI, ED, adsorption, chromatography.
-> ULVANS project
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Polysaccharides feedstocks = hydrocolloïdes from both marine and terrestrial sources
13
Feedstocks are of inegal importances Large established markets : Alginates (E400-405) : brown seaweeds and Laminaria Carraghenanes (E 407) : red seaweeds, Chondrius, Eucheuma Agar (E406) : red seaweeds, Agar-Agar, Gracillaires Pectin (E440) : apple, citrus, … Cellulose (E460) : wood, cereal straw,… Starch (E1400) : potatoe, wheat, maize, pea. No markets or lower size markets « yet » : ulvanes, xylanes, rhamnanes, mannanes, arabinogalactanes,
arabinoxylanes, fucanes, porphyran, …
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Bioactive seaweed Oligosaccharides
Brown seaweeds •Laminarine (b1-3 glucane) •Oligoalginates (D-manuronate, L-guluronate) •Fucanes (sulfated polysaccharides)
Green seaweeds •Ulvanes (sulfated polysaccharides : glucuronic acid, iduronic acid, rhamnose, xylose, glucose)
Read seaweeds •Sulfated Galactanes
14
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Origin of elicitors and stimulants? What is their impact
Traditionnals use « re-discovery » -> recent insite Technology transfert from food, healh or cosmetical domains to Agronomy Recent structured screening (physiology, proteomics) -> since the1980’s
Frequent common point with elicitors : they are inducing plant defense mechanism that are expressed at high level in the resistant varieties. A few secondary metabolites do have biocide, biostatic or direct inhibition properties on some pathogens organisms. Frequent point for stimulants : They can counteract some nutrion limitations from the soil or from the leaves.
15
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Ulva Sp : « Sea lettuce »
16
Allowed for food use : most consummed seaweed in Europe Tradionnaly harvested on the Britany costs Consummed fresh, finely chopped together with vegetables, cooked or dried High content in proteins (18 %) and polysaccharides High content in sulfur Vitamines A, B1, C Minerals (30 %) including :
– I, Ca, Cu, Si – Fe (12x lentils, = 2x wheat germ contents ), – Mg (up to 2 - 3 %)
-> ULVANS project
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Advantages / weakness of seaweeds sources
17
Seaweeds flowed on the sand by the tide: -> legal status = waste – Decomposition is started, – High microbial load, – Hich sand load, – High variability, -> heavy and costly processes. Weak and variable quality.
Sea harvested seaweeds: -> legal status = natural ressource – Fresh and well preserved – Less microbial load – Low sand pollution -> better characterized raw-material with higher quality, simple
processes and more reliable starting material.
-> Prefered source for ULVANS project
-> Good source for composting
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium 18
Seaweeds can help agriculture to lower some productivity
limitations
111
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Produce more with less - Produce better
19
stresses on the phyllosphere or on the rhizosphere are inducing productivity losses and lower crop yield
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Strategies to overcome crop plants stresses
Abiotic stresses •Genetic & plant selection • Stimulation & potentialising defense mechanisms -> seaweeds
Biotic stresses •Genetic & plant selection • Agrochemicals • Elicitors & potentialising defense mechanisms -> seaweeds • Biological pest control
Nutrition stresses • Agronomy • Fertilization •Germination stimulation -> seaweeds • Soil fertility and root development -> seaweeds
Chemical stresses • Pesticides Best-practices & more specific molecules • Formulation together with physiostimulants -> seaweeds
20
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
PRP EBV specific traits
- Mineral Inducer Process (coper, sodium, magnesium, sulfur, manganese, boron,…) - Major element: potassium
Improved response of plant to stress Rhizosphere stimulation
Improved plant growth, yield and yield components
Ulvans project will increase
theses effects
21
112
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
0
100 000
200 000
300 000
400 000
500 000
0 200 400 600 800 1000 1200
Tem ps (s)
Relative Luminescence Unit
H2O Tém oin
PRP Tém oin
H2O Chitine
H2O Chitine
PRP Chitine
PRP Chitine
H202 early signal after PRP EBV spraying
H2O2 production in response to oligo-chitosan (10 μg/ml) on young plants of Arabidopsis thaliana pre-treated with PRP EBV (2%) or with water (control)
22
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Calcic response of plant after PRP EBV spraying
Choc mécanique lié à la
manipulation
Induced calcic response early signal in Arabidopsis thaliana after PRP EBV spraying at different doses
0
0,5
1
1,5
2
2,5
0 100 200 300 400 500 600 700
Time (s)
[Ca2
+] (m
icro
M) H 2 O
0,01% 0,10% 0,50% 1,00% 2,00%
23
Olmix Algae Symposium - Sept. 10th, 2012
Root growth stimulation after foliar spraying by PRP EBV under different stress conditions
0
0,5
1
1,5
2
2,5
3
3,5
4
Accroissement racinaire (m
m)
Tém oin sur m ilieu norm al
Tém oin sur m ilieu stressant
(NaCl)
Tém oin sur m ilieu stressant
(m annitol)
PRP EBV sur m ilieu norm al
PRP EBV sur m ilieu stressant
(NaCl)
PRP EBV sur m ilieu stressant
(m annitol)
Arabidopsis thaliana root elongation 24 hours after PRP EBV spraying on leaves in different conditions with or without stresses
Olmix Algae Symposium 24
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Growth after severe hydric stress
Young plants of Arabidopsis thaliana after 10 days watering following 14 days of severe water stress.
Pre-treated with PRP EBV
Témoin
Control
25
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Profitability of foliar spraying
26
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium 27
Questions to be faced for a fertilizer producing compagny
regarding elicitors
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Regulation is not harmonized and not favourable
28
Heterogenous regulations in the E.U. countries.
E.U. regulations (2012) • Fertilizers : Yes • Amendments : No (only national) -> E.C. in 2014 • Pesticides : Yes • Elicitors & phytostimulants : No, 3 exceptions (DE,IT,ES) + « PNP » in France
Paradoxal situation • Some farmers do have their own experimental basis + some good practices • Intentions (ex : ECOPHYTO-2018 in France) • Social opinion heavy tendency towards sustainability • Scientific litterature & patents on elicitors • Large and increasing number of R&D projects
Regulation is a limiting point for players together with the registration cost
This situation favors chemical groups (already structured + critical size)
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Large offer diversity of « natural » molecules listed as elicitors or stimulant
29
Oligosaccharides -> seaweeds, vegetable, microbial, shellfish sources or sucrochemistry
Proteins -> ex : harpine, enzymes, … Aminoacides & related -> ex : L-proline, glycine bétaïne,… Lipids -> ex : brassinosteroides, phospholipides, … Metabolites -> ex : BABA, acetyl salicylique acid, ascorbique acid,
brassinostéroïdes, stilbènes, terpènes,… Plant extracts -> ex : Trigonella, Artemisia, dimers of ferulic acid, … Lignine-like -> ex : lignosulfonate, humic acids, fulviques acids, leonardite,… Microbial cell walls -> ex: yeast, bacteria, fungi. Living microorganisms -> large variety : producing elicitors, antagonists of
pathogènes, predators, nutrition promotors, mycorhizes, rhizobia,… Minerals -> phosphites, copper, silica, …
Effects are not fully understood. Interactions are not known. Which one to chose? Price/value ratio ? Which association and /or formulations are the best ?
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
What are industrials approaches for having a biological active solution?
30
Integrator : buying and formulate an already existing molecule -> Short-term (R&D is already done outside). -> Proportional cost. -> Exclusivity is diffcult to have or on short-term and limited area.
Screening available raw-material witch are not qualified for « agriculture » -> Midle term; -> Intermediate investment (R&D, formulation, agronomy), applicative IP possible; -> Risk on raw-material sourcing, variability (sometime).
Specific development of an active -> ex : ULVANS collaborative project -> Midle-long term -> significant investment -> Sourcing & quality are easier -> Exclusive, IP, homologation is possible.
115
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Projects phases for a new seaweed product development
31
Regulation & Intellectual property surveys Screening & Characterisation of raw-materials Sourcing : choice, security, traçability, quality, cost
Added value and functions : biological & functional properties
Industrial goals : product & process -> process signature
Experiments & Agronomical trials (microcosm->field)
Formulations -> Applications
Development -> Scale-up, MKT, Commercial items
Homologation : Yes / No ?
Time to market : 3-5 years for fertilizers, 5 -10 years for elicitors
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Seaweeds applications in agronomy
32
Seed coating Solid Fertilizers Ferti-irrigation Foliar spraying -> need for formulations design for each application -> need further segmentation for crop species
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Seaweeds applications
33
Seed coating •Source of nutrients for seed
- -> Germination, growth, root developement -> direct •Source of nutrients for topical microflore microflora
- -> Root microbial ecosystem - > indirect nutrition + microbial consortia composition on rhizoplan
Solid fertilizers for soil •Nutrient source for microflora (base of soil food-web). •Elements & oligoelements of marine source, solubles & bio-availables. •Active on rhizosphere ecosystem composition -> nutrition & plant health.
Foliar spraying •Nutrients for leaves •Direct signals (if elicitors are recognized by leaves receptors). • Indirect signals (if hydrolysis on phyllosphere)
116
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Interesting molecules from Ulva seaweed for agriculture
34
Polysaccharides & oligosaccharides – Source of energy, C, S & biological activity
Proteins & peptides – Source of N & biological activity
Minerals et Oligoelements – Bioavailable & soluble -> co-factors of enzymes – Naturaly complexed by ionic agropolymers (ex: carboxylic & amine functions).
Secondary metabolites and small molecules – Large number, diverse & bioactives – More documented in the health & cosmetic domains
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Stabilization
35
Solid forms – Drying (energy cost for low AW)
Liquid forms – Concentration & osmotic pressure (! Solubility !) – pH – Conserving additives – Biocides (regulation + side effects issues))
-> aging testing for stability
!!! Good molecules for plants are also good for microorganisms !!! -> need for stabilized formulations.
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Seaweeds & Industrial Quality
36
Raw-material freshness – Reproductibility (except seasonal and localizations factors) – Preserve lateral functions on polymers -> Activity – Supply chain
• Integrate : Harvest – Logistics - Process
-> cost = f(volume) + quality
Processes – Optimization of extraction-> selectivity & yield – Controlled DP et DS -> activity for ligands & receptors
Industrial limitations to be overcome !!! – sand -> abrasion ! – salt -> corrosion proof equipments
117
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Ulva efficiency
37
Classical fertilizers positive list (N, P, K, Ca, Mg, K, Na, S, B, Co, Cu, Fe, Mn, Mo, Zn). Seaweed brings some more rare and “free” elements linked to natural molecules
– Directs nutrition effects Stimulants molecules (Improve efficiency of fertilizer
elements, improve photosynthesis and major element capture from soil by rhizospheric microflora,…).
– Both directs or indirects effects Elicitors of plant defense mechanisms
– Efficiency = f (Stress type and intensity) ? – Efficiency = f (patho-system) ? – Witch mechanisms & witch messagers ? – Robustness towards genetic variability of the crop
plants and of the pathogens too ? – Is it robust towards pedo-climatic conditions in the field?
Lot of question to solve!!!
LAB to
FIELD
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
PRP SOL impact on soil microflora
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Project done with:
« Determine the impact of PRP SOL on some biological elements from the soil ecosystem : microorganisms, earthworms, plants ».
Olmix Algae Symposium - Sept. 10th, 2012
Impact of PRP SOL on soil microflora
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3 indicators where measured in a randomized microcosm experiment followed by a 3 years field experiment:
Plant biomass
Enzymatic activities in soil
Bacterial population structure by
quantitative PCR analysis
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Protocole
4 ecosystems are considered
soil* Soil +Earthworms**
Soil + plant***
Soil +Plant
+ Earthworms
* Soil : silt-clay type with no limitation (1 kg soil/ pot, no stress) ** Earthwoms: Nicodrilus giardi or Allolobophora terrestris or Aporrectodea terrestris, anecis specie (6 g/kg dry soil) *** Plant : Ray Grass (1 gramme / pot)
2 treatments / ecosystem: Control – PRP SOL
Microcosme where followed for 45 days at 20 – 23 °C, 3 pots / object
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
PCR Results
Écosystem Soil Soil
Earthwom Soil
Soil Plant
Soil Earthworm
Soil Earthworm
Plant
Soil Plant
Soil Earthworm
Plant
treatment Control control PRP SOL control PRP SOL Control PRP SOL PRP SOL 41
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Similitude dendrogram from the PCR gels
S
S+M
S+P
S
S+M
S+P
Soil
Soil + PRP SOL
Soil + plant
Soil + earthworm + plant + PRP SOL
S+E
S+E+M
S+E+P
S+P+M
S+E+M
S+E+P
S+P+M
Soil + earthworm + PRP SOL
Soil + earthworm + plant
Soil + earthworm
Soil + plant + PRP SOL
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Enzymatic Results in the ecosystem soil + plant + earthworm
0
20
40
60
80
100
Sol + plante + ver Sol + plante + ver + PRP SOL
Quantité de phénol libérée / g de sol / h
Activité de la phosphatase alcaline
0
50
100
150
Sol + plante + ver Sol + plante + ver + PRP SOL
Quantité de phénol libérée / g de sol / h
Activité de l'α-glucosidase
0
100
200
300
400
500
600
700
Sol + plante + ver Sol + plante + ver + PRP SOL
Quantité de phénol libérée / g de sol / h
Activité de la β-glucosidase
0
40
80
120
160
200
Sol + plante + ver Sol + plante + ver + PRP SOL
Quantité de phénol libérée / g de sol / h
Activité de la β-xylosidase
PRP SOL: induce significant
increase for : Alacaline phosphatase (x 3),
β-xylosidase (x 2,5), α-glucosidase (x 4), β-glucosidase (x 1,5)
(P<0.05)
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
PCA Statistical analysis
Phosphatase acide
Phosphatase alcaline
-glucosidase
b-glucosidase
N-acetylglucosaminidase
b-xylosidase
FDA
-1
1-1 1
Axe 1: 35 %
Axe 2: 21%
solsol+plante
sol+vers
sol+PRP
sol+plante+PRP
sol+vers+PRP
sol+plante+vers
sol+plante+vers+PRP
A: Correlation circle B: Projection of objets on principal axes
sol sol+plante
sol+ver
sol+PRP SOL
sol+plante+PRP SOL
sol+ver+PRP SOL
sol+plante+ver
sol+plante+ver+PRP SOL
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Olmix Algae Symposium - Sept. 10th, 2012
0,0
0,3
0,6
0,9
1,2
Tém oin PRP SOL
poids sec (mg)
Production de biom asse dans le dispositif sol + plante + vers de terre
x 2
0,0
0,4
0,8
1,2
1,6
2,0
Tém oin PRP SOL
poids sec (mg)
Production de biom asse dans le dispositif sol + plante
x 1,7
Plant biomass production
Under controlled conditions and without any stress, PRP SOL can induce a significant increase of plant biomass production.
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« The use of seaweeds in plant protection »
Adeline Picot, Plant Pathology team - Vegenov-BBV
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Outlines
1. Presentation of Vegenov
2. Elicitors: a promising strategy in plant protection
3. Seaweeds: a good source of plant defense elicitors
4. Seaweeds: additional uses in plant nutrition
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
1. Presentation of Vegenov
3
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
More than 20 years of experience on plant research and development
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Olmix Algae Symposium - Sept. 10th, 2012
Main fields of investigation
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Over more than 50 plant species
Main fields of investigation
Breeding (haplomethods, fingerprinting, marker-assisted selection and evaluation of plant disease resistance)
Plant product quality (sensory and nutritional analysis)
Crop protection (Epidemiology, Evaluation of phytosanitary products including plant defense elicitors)
- Pathogenicity assays - Biocide effects of plant products - Long-lasting and systemic effects - Study of the modes of action: biocide effect
and/or plant defense elicitor
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
2. Elicitors: a promising strategy in plant protection
6
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Wind, Rain, hail …
UV ...
Pests
Micro-organisms
Hydric stress/osmotic stress…
Nematods
Parasitic Plants
Biotic and abiotic stresses encountered by plants during their life cycle
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
How to protect plants?
8
« Classical » methods
« Alternative » methods
Genetic resistance
Conventional Phytosanitary products
Use of beneficials
Prophylaxy
Induced resistance and the use of elicitors
Plan Ecophyto : to reduce the pesticide use by 50% before 2018
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Definition of elicitors
Elicitors are molecules or non-pathogenic
microorganisms, able to induce physiological
modifications in plants, locally or systemically,
leading to the activation of plant defense
mechanisms.
9
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Particular case: priming
10
priming Infection by a pathogen
Defense mechanisms are not, or slightly,
induced
Defense mechanisms are strongly induced,
compared to untreated plants
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Wide diversity of elicitors
Wide range of molecules Ions, aa, proteins, glucids, lipids, secondary metabolites,…
Wide diversity of origins Microorganisms, plants, seaweeds, animal,… and chemical synthesis
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Perception of the elicitor
Signal Transduction
Activation of plant defense pathways
Resistance
Induction of plant defense pathways after plant treatments with elicitors
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Cell wall
Plasmic membrane
Ca2+
H+
Cl- K+
Ca2+
Oxidative burst
O2
H2O2
O2- Oxydases
1. Signal perception and transduction
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Cell wall
Plasmic membrane
Ca2+
H+
Cl- K+
Ca2+
MAPK
Signal amplification
+ + +
SA-dependent pathway
Et/JA-dependent pathway
Other defense pathways….
Enhancement of physical and chemical barriers:
- Lignins - Phytoalexins
-PR Proteins: glucanase, chitinase, …
2. Activation of plant defense pathways
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Cell wall
Plasmic membrane
Ca2+
H+
Cl- K+
Ca2+
MAPK
Signal amplification
+ + +
SA-dependent pathway
Et/JA-dependent pathway
Other defense pathways….
Infection Pathogen
Enhancement of physical and chemical barriers:
- Lignins - Phytoalexins
-PR Proteins: glucanase, chitinase, …
3. Resistance
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Advantages and limits
Advantages • Supposed to be safer than conventional pesticides due to the
indirect mode of action
• Risk of developing resistant strains is limited
• Broader spectrum of action
Limits • Treatments are preventive
• Efficiency sometimes weak and unsteady, especially under conditions of production in the field (possibly linked to formulation problems, interferences with cultural conditions such as the variety, the abiotic and biotic stresses, plant nutrition , …)
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
3. Seaweeds: a good source
for plant defense elicitor
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Seaweeds: a good source for plant defense elicitor
Treatments with seaweed extracts have been shown to enhance plant protection against a wide range of pests and diseases:
Nematodes
Fungal pathogens: Plasmopara viticola (in grapes), Sclerotinia sclerotiorum (Arabidopsis), Colletotrichum trifolii (alfalfa), …
Bacterial pathogens: Pseudomonas syringae (Arabidopsis), interference with quorum sensing…
Other pests: aphids and other sap-feeding insects (avoidance effect on plant treated with seaweed extracts)
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Seaweeds: a good source for plant defense elicitor
A variety of molecules in algal extracts can be potent elicitors of plant defense against pests & diseases:
Oligo and polysaccharides: - Agars - Carrageenans - Alginates - Laminarans - Sulfated fucans - Other complex mucilages
Antioxidant polyphenols
…
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Laminaran
Laminaria digitata
Λ-carrageen
an
Soliera robusta
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Example of seaweed extracts evaluated at Vegenov on tomato gray mold (Botrytis cinerea)
20
~ 25 days
Sowing Preventive treatment
(4-leaf-stage)
5 days
Inoculation
5-7 days
Humidity at 100%
Rating of disease severity:
necrosis length
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Protection efficiency, in % compared to water control10
0%
55%
53%52
%47
%50
%46
%
44%
0%
20%
40%
60%
80%
100%
SE1
SE2
SE3
BABA
Bion
50W
GSE
4SE
5
Rovr
al
Protection efficiency ≈ 50% with little to no impact on fungal growth and spore germination
Efficiency on tomato gray mold
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Olmix Algae Symposium - Sept. 10th, 2012
Examples of mechanisms of action
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Seaweed compounds
Pathosystem studied
Induction of plant defense
reactions References
Laminarin Tobacco/ Erwinia carotovora
PAL, LOX, SA caffeic acid, O-
methyl transferase
Klarzynski et al., 2000
Carrageenan Arabidopsis / S.
sclerotiorum Tobacco
JA-dependent response
Sangha et al., 2010;Mercier et
al. 2001
Extracts of Ascophyllum
nodosum
Arabidospis / P. syringae and S.
sclerotiorum
JA-dependent response
Subramanian et al., 2011
Extracts of A. nodosum Spinach / nematode
Phenolic components, flavonoids,…
Fan et al., 2010
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium Olmix AlgaeOlmix Algae Symposium
Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
4. Seaweeds : additional uses in plant nutrition
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Fig 1. Schematic representation of physiological effects elicited by seaweed extracts and possible mechanism(s) of bioactivity, From Khan et al., 2009 (Journal of Applied Phycology) 24
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Olmix Algae Symposium - Sept. 10th, 2012 Olmix Algae Symposium
Conclusions & Perspectives
Seaweeds: important source of nutrients and bioactive compounds, environmentally-friendly and safe, renewable bioresource
Promising results in plant protection against various plant and pest diseases
Other applications in plant nutrition as biostimulants of plant growth,…
More research is needed concerning: - the mode of actions - the identification of key gaps to their development to optimize the efficiency
in the field
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Thanks for your attention!
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Olmix SAZA du Haut du Bois
56580 Bréhan - FRANCE+33 (0)2 97 38 81 03
www.olmix.com