ADVANCES IN THE DEVELOPMENT OF FUNCTIONAL ADVANCES IN THE DEVELOPMENT OF FUNCTIONAL FOODS BASED ON RESOURCES OF ANDEAN FOODS BASED ON RESOURCES OF ANDEAN

BIODIVERSITY: THE CASES OF MACA AND YACONBIODIVERSITY: THE CASES OF MACA AND YACON

IVAN MANRIQUE AND WILLIAM ROCACIP, Lima-Perú

Module 9: “Biodiversity, Biotechnology and Intellectual Property Rights of Functional Foods”. VI ENCUENTRO LATINOAMERICANO DE BIOTECNOLOGIA AGROPECUARIA Y FORESTAL-

REDBIO 2007. October 22-26, 2007, Viña del Mar, Chile

AHIPA>95% amylopectineRotenone (insecticide)(-)

ACHIRALarge starch grainsFlour, noodles, adhesives(MEX, VNM)

Andean roots and tubers

ARRACACHASmall starch grainsVit, A, C; Baby foods(BRA)

MASHUAGlucosinolates, anthocyaninsNematicide, insecticideAnti-inflamat.(-)

MAUKAProteins(-)

OCAOxalatesPolyphenolsCarotenoids(USA, NZL, CHN)

OLLUCOPigments(DEN)

NATIVE POTATOESPolyphenolsCarotenoids, anthocyaninsVit. C; Ca(-)

MACAGlucosinolatesAlkaloids, phytosterolsUnsat. fatty acids(-)

YACONFructooligosaccharides (FOS)Phenolic compounds(JPN, NZL, KOR, CHN, BRA, PHL, CZR)

Expected benefits from adding value (*) to Andean root and tuber crops

• Improved food security and health (combat ‘hidden hunger’)(**)

• Increased income, thereby contribution to poverty reduction

• Shared benefits to stakeholders along the value chain: employment generation

• Increased incentives to maintain biodiversity-cultural diversity

• Enhanced access to biodiversity for wider utilization and livelihood functions

(*) ADDING VALUE : connecting a resource to a use through identifying the bio-chemical and genetic bases of phenotipic traits, (**) GFU/UPS (2003)

Maca (Lepidium meyenii Walp .) Brasicaceae

• 1990. only 50 ha planted-declared under extinction risk; compositional studies start• 1995. Swiss-chemical labs, Peru (with CIP): research & outreach• End 90’s: demand of maca flour grows in urban markets

Major functions: altitude-related infertility alleviation, sexual enhancement in mammals, vitality promotion in men & womenInternet publicity as “Andean Viagra”: maca exports increase

• 2006. US$ 4 million due to 500 MT maca flour exports, 1500 farm families involved (6-7 thousand jobs)

• Pre-Inca & Inca times: foods and rituals, warriors’ diet, tribute: 15 MT to Cusco

• Conquest & Colonial times: animals recover fertility, tribute (100 yrs)

• Post-colonial knowledge / use: energetic and reconstituent

• Recent times:

Traditional use• Boiled and roasted: soups• Dried (stored): drinks (juice, cocktails), desserts

Nutritional value (**)

Fresh: + 80% waterDry matter: Carbohydrates Protein Essential aa (lysine, arginine) Unsaturated fatty acids (linoleic, oleic, lauric, palmitic) Fiber Minerals (Ca, Fe)

Maca crop features I

(*) CIP (2005-06)

Crop distribution (*) • Genus Lepidium : 175 species ; Andes : 14 species• Maca : possibly only cultivated species

• Central Andes PER: 90 - 95% • Altiplano PER-BOL, other: remaining %

(**) Dini et al (1994); Lock et al (2002)

• Glucosinolates(*)

GlucotropaeolinSinalbineBenzylglucosinolateHydroxy-benzylglucosinolateMethoxy-bencylglucosinolate

• Alkaloids(**)

MacaeneMacamide

• Flavonoids

• Phytosterols(***)

CampesterolStigmasterolSitosterol

Maca functionality

GLS

IsothiocyanatesThiocyanatesNitriles, etc

• Plant defense• Carcinogenic detoxification• Block cancer cells

proliferation (****)

(*) Marthe et al (2002), (**) Piacente et al (2002), (***) Kyeong-Jun et al (2005) ; Norato et al (2003)

Myrosinase

•Energy reconstitution•Fertility recovery

Maca crop features II

• A bi-annual species: vegetative & reproductive phases• Extreme environment conditions ( > 4,000 masl, strong winds, low RH, -4 to 15oC)• Variable productivity: 1-10 MT / ha• Depletes soil nutrients quickly (long rotation periods, high organic content)• Narrow intra-species variability:

Autogamous (cleistogamy)Octoploid (2n=8x=64)Strong GxE: yield, GLS content, etc (*)

Yield (Cerro de Pasco)(**) ++++++ ++ ++++Glucosinolates (umol/g)(**) 13.3 17.9 10.8Antioxidant capacity (ugTE/g)(**) 7500 11400 9800Rats’ prostate size reduction(****) - + -Men’s spermatogenesis increase(****) - - +

Parameters Yellow Red Black

(*) Locher. Thesis; CIP-ETH collaboration (2006). (**) Clément et al. Poster; work with CIP (2007)(***) Gonzales et al (2005). (****) Gonzales et al (2001)

:

Year FOB value (US$)

Volumen (Kg) Price unit (US$)

1999 2.2 million 485 thousand 12

2000 1.4 95 152001 1.7 189 92002 2.8 313 8.82003 3.3 308 10.82004 3.4 405 8.52005 3.7 417 8.82006 3.8 497 7.6Total 22.3 2,709 80.5

Per year 2.8 338.6 10.1

Exports 1999-2006

From 2003, volume of exported maca flour increased and unit price decreased

Maca productFOB value 2006 (US$) Volume (kg) Price unit (US$)

Flour 1.6 million 274 thousand 6Extract 0.7 18 39Capsules 0.5 20 23Gelatinized flour 0.4 44 8Micropulverizada flour 0.1 18 6Maca candy 0.1 16 4Other presentations 0.5 108 5Total (7 products) 3.9 498 91Average per product 0.56 71.1 13

80% of exports consist of maca flour

Added value to export maca products

Maca: export market trends (*)

(*) Biocomercio, Peru. http://www.biocomercioperu.org/estadisticas.htm

In 2006, out of 50 enterprises involved, 5 obtained 35% of the total export value : US$ 266,000 each of 5 –vs- US$83,000 each of remaining 30

Patent: a maca extract for pharmacological applications (US patent # 6,267,995; 2001)

Case “Pure World Botanicals”-”Naturex”

Extract composition Claimed application

5-9% bencyl-isothiocianates Cancer

1-3 % sterols Sexual disfunction

20-30% fatty acids

10% or more macamidesAlso patented: process for obtaining the above extract

The Pure World Botanicals’ patent utilized basic principles of traditional process for preparation of the extract (Yllesca 1994). “Naturex” recent offer (2007):Grant free patent licenses to Peruvian-owned companies to manufacture and sale maca extract. Naturex foundation will make development grants (education, medicine, and basic needs) to communities , consistent with the principles of the CBD.

Higher levels of maca valorization

IP status referred to the maca patent:

$ 2.0

Dry maca

$ 0.4/kg

Fresh maca

$ 6.0

Maca flour

$ 8.3

Gelatinized flour

$ 23.1

Maca capsules

$ 39.0

Maca extracts

Genes MoleculesBioactive

compounds

Levels of maca added value

FARMERS’ FIELDSPROCESSING

PROCESSING

Challenge: equitable distribution of benefits

Colombia

Ecuador

Perú

Bolivia

Argentina

Geographic distribution

Inter-Andean valleys and temperate low lands (1800-3500 masl)

Yacon (Smallanthus sonchifolius). Asteraceae

•Old registers: Candelaria Culture, Argentina (1000 AD) and Nasca Culture, Peru( 500-1200 AD)

Smallanthus : 21 species ; all in the Americas

Yacon : allo-octoploid 6A + 2B ; 2n = 8X = 58 ; A =7 , B = 8 (*)

Low seed set and low seed viability (**)

Peru : 6 collections = 399 accessions

(*) Ishiki et al(1997)(**) “Sarada Otome “ –first yacon variety using embryo rescue , Japan.

Ecuador (1982)

New Zealand

Brazil (after 1989)

Japan (1985)

RussiaCzech Republic

China Taiwan

KoreaUnited States

Philippines

• Tradicionally, the yacon crop was restricted to subsistence farming• Market interest: when yacón leaves the Andean region and first scientific

research is reported (1989, Japan)

Yacon migration

Antioxidant

Fructooligosaccharides (FOS)

Anti-glycemic

50 – 70 % FOS (Total CHs)Content: range 2-12 g/100 g (*)

(*) CIP (2006): 8 accessions of CIP germplasm bank; (1) Pedreschi et al 2003; (2) Genta et al 2005; (3) Lobo et al 2007; (4) Valentova et al 2004; (5) Aybar et al 2001

Chlorogenic acidCaffeic acid and derivatives

Yacon functional value

Yacon functionality

• FOS: low caloric (1-1.5 kcal/g)• FOS: promotes colon beneficial bacteria (1) • FOS: decreases blood triglyceride level (rats) (2)

• FOS : increases Ca, Mg absorption; Ca bone accumulation (rats) (3)

• Leaf extract reduces liver glucose (diabetic rats) (4)

• Leaf infusion reduces blood sugar level; increases insuline (diabetic rats) (5)

As export volume increases, unit price goes down

Biocomercio, Peru. http://www.biocomercioperu.org/estadisticas.htm

0

50,000

100,000

150,000

200,000

250,000

300,000

2001 2002 2003 2004 2005 2006

FOB value (US$)

Unit value (US$/kg)

Volume (kg)

13.1 US$

2.7 US$

30 thousand US$

251 thousand US$

2 thousand kg

93 thousand kg+700%

-80%

+4500%

Yacon export market trends; products: syrup, leaflets, flour, tee

Yacon: research opportunities

1. Variation in key traits (Manrique 2006, CIP)

2. Yacon clinical studies with humans (***)Yacon syrup effects on: triglyceride, glucose and insuline blood levels; Ca and Mg absorption; constipation; toxicity

3. Yacon FOS degradation after root harvest

(***) Collaborative work: Univ,. Tucuman, Univ. Basilea, CIP

FOS (%DM) 44 – 63 12.6 – 55.0Root DM (%) 12 – 14 6.0 – 18.4Root yield (t/ha) 37 – 57 2.8 – 49.7Foliage yield (t/ha) 10 – 13 0.5 – 11.7Precocity (days) 180 – 240 198 – 316

Range genotypes(*) GxE (8x6)(**)

(*) Highest yields, precocity: AKW5075, resistence to oxidation: CLLUNC118(**) Highest yields, precocity: OXA2, highest DM: CAJ1

Approaching yacon FOS degradation

CH2OH

2 5

3 4 6

CH21

CH2OH

2 5

3 4 6

CH2OH1

CH2OH

1 CH2

CH2OH

2 5

3 4 6

G F

F

F

F

CH2OH

2 5

3 4 6

CH2OH1

F(2 - 10)

FOS

Fructan fructosil transferasa (FFT)

Fructan hidrolasa (FH)

• FOS degrades to simple sugars after harvest

• After 10 days of storage, 40% FOS has degraded to F, S and G(*)

• Cut down FH activity at harvest

• Isolate and clone genes involved in FOS biosynthesis (SST y FFT), and express in yacon foliage and/or other biological systems

(*) Graefe et al 2003 (CIP). Field Crops Research 86(2-3): 157-165

$ 0.2 / kg

Fresh roots

$ 0.3

Selected roots

$ 8.4

Syrup

$ 8.8

Dehydrated leaves (tee)

$ 17.7

Capsules

$$$

FOS

Genes MoleculesBioactive

compounds

Levels of yacon added value

Challenge: equitable distribution of benefits

FARMERS’ FIELDSPROCESSING

PROCESSING .

COLLECTIONSEx-situ

AGROBIODIVERSITY•Cultivated•Wild

TRADITIONALKNOWLEDGE

Whole plantTissues/Cells

extraction

DNA extraction

Chemical extraction

MORPHO-AGRON.CHARACTERIZ.

Profiling

GENETICCHARACTERIZ.

Profiling

BIOCHEMICALCHARACTERIZ.

Profiling

Traditional breedingTissue culture (HB)MAS Genetic engineering

GENETICALLY IMPROVED

GERMPLASM

SELECTED GERMPLASM

SELECTED GERMPLASM

Extracts Chemical libraries

Functional bioactive

compounds

Molecules

Genetic maps

Genetic markers

DNA libraries

Gene libraries

Genes

(*)

(*)

(*)

(*) Phenotypic, genotypic assays; Contents; Bioavaiability; Toxicity; GxE ; Maca : Arabidopsis genetics and genomics

Adding value to R&T crop biodiversity through biotechnology

Genomics *

Metabolomics*

                                                                                                                                             

GRACIAS!GRACIAS!Acknowledgments:Acknowledgments:

Carlos Arbizu, CIP, LimaCarlos Arbizu, CIP, Lima

Alberto Salas, CIP, LimaAlberto Salas, CIP, Lima

Cinthya Zorrilla, CIP, LimaCinthya Zorrilla, CIP, Lima

Rommel Aguilar, UNDAC, Cerro de PascoRommel Aguilar, UNDAC, Cerro de Pasco

David Ponce, UNDAC, Cerro de PascoDavid Ponce, UNDAC, Cerro de Pasco

Michael Hermann, BioversityMichael Hermann, Bioversity