Building our Biobased Economy together

BE-Basic FoundationAnnual Report 2015

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ECO BIOThe first edition of theECO-BIO Conference

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InternationalalliancesSeizing opportunities

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Facts and figures

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Activities

Foreward

BE-BasicFoundation GovernanceAccountability

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On the thresholdof transition

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Flagship 1Second generation carbon-basedcompounds 12Flagship 2Nitrogen-based specialities 14Flagship 3Sustainable soil managementand upstream processing 16Flagship 4Bioconstruction materials 18Flagship 5Microbial production of biofuelsand biorenewables 19Flagship 6Synthetic biology 20Flagship 7High-throughput experimentationand metagenomic mining 22Flagship 8Environmental impact of chemicals,biobased molecules and processes 24Flagship 9 & 11Societal Embedding, EBD Programme 26Flagship 10Genomics for industrial fermentation 30Flagship 12Iso-Butanol Platform Rotterdam (IBPR) 32

ChairDr. Herman van WechemIndependent consultant AlternativeTechnologies and Climate Change issues

Drs. Sytze KeuningChief Executive Officer, Bioclear

Dr. ir. Jan van Breugel Independent BioBased Innovationconsultant

Prof. ir. Karel Luyben Rector Magnificus, Delft University ofTechnology

Prof. dr. ir. Louise Vet Director, NIOO-KNAW

Dr. Manfred KircherChair Advisory Board, CLIB2021 ClusterIndustrielle Biotechnologie

Dr. ir. Kees de GooijerChief Inspiration Officer for TKIAgri&Food, TKI Biobased Economy

BE-Basic Supervisory Board IPRC panel composition

The IPRC members involved in the evaluation process of the BE-Basic programme are:

Prof. dr. Urs von StockarEcole Polytechnique Fédérale de Lausanne, Switzerland

Prof. dr. Matthias H.C. ReussUniversity of Stuttgart, Germany

Prof. dr. Sven Panke Department of BiosystemsScience and Engineering, ETH Zurich, Switzerland

Prof. dr. Martin E. FederUniversity of Chicago, USA

Prof. dr. Rik Eggen Eawag, Dübendorf, Switzerland

ColophonInterviews: Mirjam de Graaf and Edith Vos (Tekstkeuken), Robin Pascoe, Trudy van der Wees

Translation: Zuidas Text and Translations

Coordinator: Sandra Ransdorp and Iva van der Voort-Polla (BE-Basic Foundation)

Graphic Design: VastinVorm

Photography: Corrie van den Berg, Henk Houtzager, Martien de Man, Anne de Jong, Verse Beeldwaren,

Mirjam de Graaf, BE-Basic

Courtesy of: Lucien Joppen, Jelle Vaartjes

We thank all scientists and parties involved for their cooperation.

BE-Basic FoundationAnnual Report 2015

20052015

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Of course we also encountered our first seriouschallenges, which were both essential and expected forthe growing maturity of the biobased sector. Low fossilprices challenged business models and technologyproblems at the new plants caused by working 24/7 withreal feedstocks (sand, mud, stones and metals) needed tobe solved. With respect to the latter, we have come torealise that treating crop residues (stover, bagasse etc) aswaste is too simplistic and harvesting methods have to bere-thought. And last but not least, the debate aroundbiomass sustainability saw a number of new peaks viainterventions in some media and a few professionalorganisations. Although frustrating at times, the publicattention shows that biobased development is maturingand beginning to provide a serious alternative to our stillpredominantly fossil fuel-based economy.

All of this shows BE-Basic is entering a new phase in itsdevelopment. Project teams have completed many (butnot all!!) of their laboratory proof-of-concepts andpiloting. Demonstration plants such as ‘our’ BioprocessPilot Facility in Delft and others are now fully operational.BE-Basic has committed its last FES budgets to thesedevelopments and so have companion programmes atEuropean and regional levels. This is why BE-Basic ishosting BioPort Holland, the coordination body of theaviation sector and why we were invited to take part inbiobased implementation projects such as the BrazilianAgropolo programme. BE-Basic has also contributed toimportant strategy documents such as the Dutch AviationVision, Action and Implementation plans, and toambitious global sustainability studies1 about biomassavailability, income distribution and job creation,biodiversity and climate impacts.

All signs show that fossil fading-out has started infinancial terms and at high rates that are difficult tomatch with investments in renewables (the ‘carbonbubble’). BE-Basic also changed its communication fromrelatively distant and neutral towards speaking-up in aninsisting and even stronger tone. Change is urgent andall financial as well as human resources need to be

focused on investing in a cleaner, more beneficial andprofitable world. BE-Basic cannot afford to sit on theside-line and must pro-actively communicate theadvantages of our integral strategies such as in the HIPprogramme, the REDEFINERY, the Dutch fuels mix andthe like. If we don’t, who will?

As you read this public summary of the BE-Basic annualreport, you will find this sense of urgency stressed by allinterviewed partners. More products and services areinvading the commercial domain but the rate of changeand implementation needs to be higher. Please join us onthis journey in 2016 towards a sustainable future andbeyond – we will get only one chance!

Bram Brouwer and Luuk van der WielenBoard of Directors of BE-Basic Foundation

1) Such as the Macroeconomic Outlook, SCOPE Report on Bioenergy and

Sustainability, Biomass studies of Cie Corbey and others.

Foreward2015 was clearly a year of change. The first large-scale lignocellulosic ethanol plants came online, a range of biobasedbuilding blocks (lactic acid, succinic acid, FDCA, itaconic acid etc) had their first applications and various biobasedfeedstocks and products have been shown to have environmental and safety characteristics which are in many casesbetter than their conventional (often fossil) counterparts. Sustainable biofuels are now broadly accepted in sectorswithout or with only limited alternatives, such as in aviation and maritime applications. And serious implementationstrategies are being embraced by the Netherlands’ national and regional governments and other authorities.

GovernanceThe executive board consists of Bram Brouwer (managingdirector) and Luuk van der Wielen (chairman). Theboard’s task is to ensure cohesion across the entire BE-Basic programme and to take care of day-to-daybusiness including decisions about funding and thecontinuation of projects. The board presents its decisionsto the supervisory board for its recommendations. Thesupervisory board monitors the actions taken by theexecutive board and represents the interests of the BE-Basic partners. In addition, all partner organisationsare represented in the Consortium Partners Assembly(CPA). The CPA offers important, non-binding advice tothe board.

The actual research takes place in Flagships, each ofwhich focuses on a specific aspect of the biobasedeconomy – from technology development to publicacceptance. In total, over 80 projects are ongoing underthe auspices of the Flagships. In addition, the BE-BasicInnovation Centre (BE-BIC) stimulates innovative start-upsand entrepreneurship in the field of the biobasedeconomy. Progress in the Flagships and BE-BIC ismonitored by the management team, in which allFlagship managers and the board are represented.

BE-Basic also hosts the Dutch EBD (Economy, Policy andSustainability) programme of TKI-BBE – the TopConsortium for Knowledge and Innovation, BiobasedEconomy, which is supported by the industry andacademic parties as well as the government (TopsectorEnergy). The EBD programme in Flagship 11 focuses ondeveloping policy to stimulate support for the biobasedeconomy among stakeholders and the general public andis closely related to Flagship 9.

In addition several TKI-BBE projects are governed by theBE-Basic Programme, such as the AMBIC project onmicrobial-based industrial production of chemicals andbiofuels and ISIM on improved stability and productivityof microbes for industrial production. Moreover, the BE-Basic Foundation hosts the EFRO-funded project

Isobutanol Platform Rotterdam (IBPR), aiming at pilotscale and industrial production of isobutanol and relatedchemicals and fuels from biomass.

AccountabilityBE-Basic accounts for its results and research to severaldifferent bodies.

FinancialBE-Basic’s programmes are funded by industrial andacademic partners as well as the Dutch government’s FESfund and other resources (TKI-BBE, EFRO). TheNetherlands Enterprise Agency (RVO.nl) monitorsresearch progress and financial accountability on thebasis of deliverables and key performance indicators(such as published papers, the implementation of newmethodology by industry and patent applications). The supervisory board has assigned an auditingcommittee to check the annual accounts of thefoundation.

Review and evaluationAll new project proposals are subject to an internal (bythe board and management) and external review by theInternational Peer Review Committee, an independentbody of external, distinguished specialists from academeand industrial leaders in the field from the Netherlandsand abroad.The BE-Basic programme is also reviewed by the IPRC onan annual basis. They discuss progress in all projects withthe Flagship leaders and draw up a confidentialevaluation report.

To stimulate innovation from within the consortium, theInnovation Evaluation Committee reviews all papers priorto their publication on possible protection of foregroundknowledge.

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BE-Basic FoundationThe BE-Basic Foundation coordinates and stimulates RD&I programmes for science and technology development,especially - but not exclusively - in the fields of environmental and industrial biotechnology. The focus is onintegral and sustainable biobased solutions that balance and optimise economic value and climate impact fornon-energetic (chemicals, materials, food & feed) and energetic (transportation fuels, power/heat) uses. Thosebalances are reached best by applying cascading and integral biorefinery concepts, and prioritise energetic usesfor those sectors without alternatives (aviation, marine and heavy road transport).

Open to new ideasBrouwer and Van der Wielen look at those hurdles as somany positive new challenges. “As we move from conceptto application, we learn new things all the time,” Van derWielen say. “We had a clear view of that spot on thehorizon, but we didn’t know how to get there. The strengthof BE-Basic is that we are always open to new ideas.” “Thatopenness is also down to the unique partnership betweenscientists, technicians, companies and governments,” addsBrouwer. “Over the years relationships of trust have beenbuilt which have set off a process of cross-pollination thathas proved enormously beneficial. It’s now becoming clearhow successful those alliances have become.”

ImplementationA growing number of techniques and biobased solutions areready to be marketed. Van der Wielen: “BE-Basic is at theforefront of biobased research and infrastructure. At ourBioprocess Pilot Facility the number of industrial requestsand indications of interest has been increasing, and ourapproach is regarded as successful internationally as well. Ifwe want to build on this success we need to be pro-active.”Brouwer: “The opportunities are there, we have thetechnology and we have the people. Much depends on theinvestment climate and that is not something we havecomplete control over. It’s an important aspect, one that wehave to tackle right now.”

PartnershipsThe transition will require a public-private approach for theforeseeable future. “We need to recognise that we are on thethreshold of great changes,” Van der Wielen said. “And thelaws of transition decree that with transition comes a certainchaos. Investors feel the risk is too big at the moment,increasingly of fossil and less and less in renewables.Governments have a role in mitigating those risks. They cando this by investing but first and foremost they can create theright conditions.” At the same time, biobased solutions haveto be supported in order to create a level playing field with,for instance, solar energy and wind power. “I sometimes jokeabout the need for a ‘state bio refinery’ in Rotterdam,” Vander Wielen says. “By that I don’t mean it should be 100%state owned, on the contrary I would be fine with just private

funding for future projects and so would the BE-Basicpartners. But we have to work together to find investors whoare prepared to consider the medium and long termpossibilities.” For Van der Wielen the long term financialsuccess of biobased solutions is not in doubt. “You only haveto look at the pension funds. The investments that they hadto write off were mainly fossil fuel related. With aninvestment of 10% of that loss we could have the wholeDutch aviation sector running on biofuel, and that is thefuture.”

Give a boost The BE-Basic biobased perspective is firmly linked to thepresent-day situation. Van der Wielen: “The importance ofthe port of Rotterdam as a transport hub is widelyacknowledged. The Netherlands is a rich country and bytaking pro-active measures to ensure a clean economy it willremain a rich country. The Macroeconomic Outlook (see page29) outlines scenarios for an industrial future based partly onthe use of biofuel embedded in the existing infrastructure. BE-Basic wants to accelerate that process. It’s one of the reasonswe are involved in Bioport Holland. It has a clearly definedlimited time path which will lend a sense of urgency to theimprovements that are needed with regard to, for instance,sustainable feedstocks, and which will boost the developmentof the product until it can stand alone in the market.”

Make choicesMeanwhile, that small spot on the horizon has been growingquite substantially. It’s a different perspective which Brouwerand Van der Wielen feel has practical implications. “There isnow a continuous pressure to forge ahead with theintroduction to market of the latest biobased solutions,” theysay. “BE-Basic will continue to focus its efforts on themovement towards implementation. We need to makechoices, we can’t research absolutely everything andanything. ‘Benefit scope’ must be at the heart of everydecision. It’s a complex matter, and there is no single rightanswer. But at some point we have to come up with the bestdecision for the time and implement it. BE-Basic is unique inits public-private partnership expertise and is happy toprovide the platform for a relevant and deliberateddiscussion on how to go about this.”

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On the threshold of transitionOnce the transition to a sustainable, biobased society seemed a distant spot on the horizon, now the momentum forchange is almost palpable. It is no longer a question of if but of when and how quickly it can be done. BE-Basic’s managingdirectors Bram Brouwer and Luuk van der Wielen have faith in the technological possibilities, but at the same time theyfeel that now is the time to act. “There is a window of opportunity but that window will not be there for ever. Noteveryone is aware of that.”

“Upscaling and improving are today’s key words,” says Brouwer. “Many solutions have reached the demo phase oreven the marketplace and that has shown us where the bottlenecks and loopholes are. It’s an important part of theprocess. Some things can’t be solved with the existing technology. With the knowledge and experience we gainalong the way, we can concentrate on finding alternative solutions. Once we take those final hurdles, we are readyfor the marketplace.”

ECO-BIOIn 2015 BE-Basic and Elsevier took up the organisation of the first edition of the ECO-BIO Conference, which tookplace from March 6 to 9, 2016 at the World Trade Center in Rotterdam. The conference provided an internationalplatform for industries, scientists, NGOs and governments to discuss the needs of a biobased economy fromacademic, environmental and societal viewpoints.

During the conference it became apparent that there areeconomical, societal and technological challenges whichneed to be solved to achieve definitive breakthroughs fora biobased economy. Delegates also agreed that the useof biomass for biofuels, chemicals and polymersproduction will be instrumental in reaching CO2-emissiontargets (COP21, Paris 2015). The biobased economy, theyargued, makes sense from an economic point of view inthe long term.

Marcel Wubbolts, chief technology officer at DSM,stressed the importance of using biomass in reducinggreenhouse gas emissions and slowing down globalwarming. “Given the increase in the world populationand the rise of the middle classes in emerging economies,we need to develop renewable energy, not only wind andsolar, but also biomass for energy andchemicals/materials,” he said. “At DSM we havedeveloped several biobased materials which offer uniqueselling points in terms of functionality and theirenvironmental footprint.”

The rise of advanced biofuels Carlos Henrique de Brito Cruz, Scientific Director at FAPESP,stressed that biomass is needed just as much as wind andsolar energy and said all three should have beenmentioned at the COP21 in Paris. “The energy share ofbiomass worldwide is hovering around the 10% mark,most of it being first generation biomass, such as sugarcane,” he pointed out. “Currently, the share of advanced- 2nd generation - biofuels is relatively low. However, thisshare is expected to rise to 70% of the total biomassvolume for energy purposes by 2035.” The shift towardsdeveloping biofuels from non-edible biomass will alsohave an impact on the sustainability discussion.According to De Brito Cruz, using 2nd generation biomassdoes not exclude discussions on sustainability issues, suchas water and land use, land rights and so on. “We needto develop crops which are less dependent on water andthat have higher yields,” he said.

During the conference, several speakers stressed thatproducts which are able to offer other functionalities donot necessarily compete on price with fossil-basedproducts. Unfortunately, the producers of one-on-onereplacements, so-called drop-in's, are experiencingdifficult times because crude oil prices are hoveringaround the 30-40 dollar level. Therefore, incentives needto be put in place to level the playing field, such ascarbon taxing or fiscal measures. This means, speakerssaid, that firm policies need to be put in place over alonger period of time. Without this, CO2 reductiontargets will not be met, and this will lead to ecologicalproblems and substantial costs in mitigating the effects ofclimate change. Furthermore, it will setback reviving localeconomies, which are highly dependent on local biomass.

New business modelsCreating new value chains, redefining value. Thetransition towards a biobased economy requires morethan just developing and marketing products, accordingto DSM’s Marcel Wubbolts. “In essence, it is aboutconnecting the renewable (biological) cycle with the non-renewable (technological) cycle, in which renewables increasingly compensate for losses in thetechnological cycle.”

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Participants 380Countries represented 40Plenary speakers 5 Speakers 22Posters over 180Company tours 6

Second generation carbon-based compoundsThe Flagship Second generation carbon-based compounds aims to develop clean, efficient and otherwisesustainable industrial processes for carbon-based chemical building blocks mainly for the chemicals,materials and fuels industries either by fermentative production or by modification of substrates orprecursors. The research focuses on the conversion of lignocellulosic materials and other biobasedfeedstocks, including their contaminants, into relevant products.

The project ‘Process development for the production ofC3-Acids from ligno-cellulosic feedstocks’ has alreadyresulted in six academic publications, and four more arecurrently awaiting approval. Wageningen University hasawarded two doctorates on the subject and a third PhDstudent is still researching new strains of lactic acid-producing bacteria. In addition, a patent has beenrequested for a new discovery making the bacteria whichproduce lactic acid more accessible to geneticimprovements. “Things are beginning to take shape,”says Mirjam Kabel.

Sugarcane bagasseKabel’s team has researched how to improve thesugarcane bagasse conversion into sugars, whichcontributes via lactic acid to the production of poly-lacticacid (PLA), a bioplastic. Bagasse, the fibrous material leftafter the juice has been extracted from sugarcane, islargely comprised of cellulose and hemicellulose but alsocontains lignin. Kabel’s team researched which factorsplay a role in the degradation of bagasse. To a morelimited extent, they also researched the role and structureof lignin. Lignin is next to cellulose the most prevalentorganic matter on earth, so it may play an important rolein the production of sustainable materials. Furtherresearch is still needed to better understand its complexchemical structure and what happens when it is brokendown. Once that is understood, the number ofapplications for lignin will be much greater.

Alkaline processA lot of time has been invested in finding a good way toturn the lignocellulosic biomass into the fermentable

sugars that are key for the creation of lactic acid. “At themoment, the state of the art approach involves diluteacid (slightly acidic, high temperature) and hydrothermalpre-treatments (high temperature without the addition ofchemicals),” says Kabel. “We have looked into whetheran alkaline process with a high pH and relatively lowtemperatures would also be an option. One advantage ofan alkaline-based process is that the lignin is partiallybroken down and can be separated from the celluloseand hemicellulose. This lignin fraction could be used, forexample, to provide the energy needed to carry out theprocess. Another advantage of the lignin separation isthat the conversion of (hemi-)cellulose to sugars proceedsmuch more smoothly. The presence of lignin slows theenzymatic conversion.” There are also disadvantages.“We have been working with sodium hydroxide which isexpensive,” says Kabel. “It is only cost effective if you canreuse it and that is difficult. Sodium carbonate wouldappear to be a good alternative even if the pH is a littlelower.”

New enzymeKabel is extremely pleased about the results. “We betterunderstand which enzymes are needed on top of theexisting cocktails. We know xylan-degrading enzymes arevery important as well as cellulases. We’ve characterizeda new enzyme – alpha-glucuronidase, which cancontribute to a better conversion. Finally, the research hasgiven us better insight into which components arereleased alongside sugars when sugarcane bagasse ispretreated and what impact these substances have onthe fermentation of lactic acid.”

‘Sugarcane bagasse: a promising feedstock for bioplastics'

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Bioplastics, the sustainable alternative to conventional plastics, are made from sugars extracted from vegetablebiomass. Can these sugars also be obtained from plants or parts of plants that are not fit for humanconsumption? One project within Flagship 1, led by Mirjam Kabel of Wageningen University in cooperation withseveral other groups of Wageningen University and Research, DSM and Corbion, is looking at the potential ofsugarcane bagasse.

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Nitrogen-based specialtiesThe Flagship Nitrogen-based specialties develops novel technologies for the production of nitrogen-containing compounds from renewable feedstocks through the use of advanced engineering ofmicro-organisms. Research within this flagship is focused on the design and optimisation of pathways forthe production of compounds that can be used as pharmaceuticals or as building blocks for materials. In addition, this flagship includes high-throughput bio-pharmaceutical process development aiming at theoptimisation of protein production.

“Say you have a protein-based product and you want toknow how one can produce it on a large scale, but youonly have a small quantity available,” says Marcel Ottensfrom Delft University of Technology. “This means youdon’t have enough material to perform manyexperiments. However, we now have the technology inhouse to carry out these experiments more efficiently andmore quickly. This research project is about carrying outmany experiments with a very small volume of protein.”Ottens’ team has looked at how process developmentcan be done more quickly (high-throughput processdevelopment) using the tiniest amounts of availableprotein. Together with the University of Twente, theKarlsruhe Institute of Technology (KIT), DSM andSynthon, they have developed a new approach to processdesign. “This approach is a combination of robotics,which generates a lot of data, and mechanistic models,which can predict the behavior of proteins,” says Ottens.“By linking these two aspects, you can experiment moreefficiently. You already have an idea of where you needto focus.” In order to automate the process, the team isdeveloping a special lab equipped with robots. Therobots may be two metres wide, but the proteins are ofnano format. It is now, however, possible to carry outsome parts of the process via a lab-on-a-chip device.

CommercialisationThe approaches taken by Ottens’ team will allow newmedicines to be developed and hit the market morequickly. “The technology we have developed is alsoavailable to other parties,” Ottens says. “An integral partof this research project was to find out if we couldcommercialise the results. We’ve taken the first steps.

A business plan has been put together for a startup toprovide this service to interested parties. There is a lot ofinterest from bio pharmaceutical firms, but it takes moreto create a successful and independent company. That issomething to look at next.” One important part ofcommercial success is ownership of the discoveries, and apatent is currently pending. “It’s just a matter of time,”says Ottens. “Patents strengthen the position of acompany. We can offer the software and infrastructurewe’ve developed to clients.”

International reachInternational companies are very interested as well in thisapproach, which Ottens describes as 'trendsetting'. “TU Delft has a name in terms of high-throughputprocess development,” he points out. “There is a reasonwhy some of the researchers who have worked on thisproject have found jobs with international companies likeNovartis and Novo Nordisk. Within BE-Basic we are partof an international consortium with important partners.For example, we have worked with a German lab that ishighly experienced in robotics. By joining forces, we areable to make major advances.”

Targeted experiments with a lab-on-a-chip

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Medicines used in the treatment of cancer are very often based on proteins. Genetic technology enablesmammalian cells to be modified so they produce a tiny amount of human protein. But how do you know whatis needed to produce that one specific protein-based drug in large quantities? The answer lies in experimentation,which can be extremely time consuming. The Flagship 2 project Peptides and Proteins: High-Throughput Bio-Pharmaceutical Process Development will enable this to be done more quickly in the future.

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Sustainable soil management and upstream processingThe Flagship Sustainable soil management and upstream processing focuses on developing and testing scenariosfor waste management from the perspective of closing cycles and the prevention of negative side effects derivingfrom a biobased economy (spilling=spoiling). The aim is to prevent potential negative side effects associated witha more biobased economy and to ensure that a biobased economy really results in a more sustainable planet.

The focus of this project is on the production of value addedproducts in key regions in Brazil using different localfeedstocks in order to draw up specific, local, sustainablesolutions. The first phase of the project involved developingthe methodology for a fully integral sustainability analysis ofentire value chains, from biomass production to logistics,(bio)chemical conversion and waste management, as well associal quality, and human and environmental health.

Total value chainOnce the most promising sustainable productionpathways have been identified, the team carries outdetailed due diligence reports for industrial piloting anddemonstration projects. These also include an analysis ofhazards, opportunities and benefits for local farmers andcommunities. “There are so many aspects to consider, andthe interesting thing about this project is that we look atthe total value chain,” says professor Patricia Osseweijerof Delft University of Technology. “It is a project whichbrings together different partners and different Flagshipsand tries to assess the impact of biofuel production acrossall aspects of local communities.”

Crops and public perceptionTake, for example, the use of sugar cane to develop jetbiofuel. “You can make jet biofuel from sugar cane, fromsoy and from lignocellulosic feedstocks, but it has beendifficult to determine which is the most interesting crop,”she points out. “If you use sugar cane, for example, youhave a crop with quite a high yield per hectare, but theprocess to make fuels is quite different from oil based crops.Public perceptions also play a role, as well as local transportissues and employment. Palm oil, for example, is the mostproductive oil crop per hectare, but has a negative image.You also need to look at maintaining soil quality, recyclingoptions for nutrients, and how you dispose of the waste.

There are so many factors attached to each crop, and noanswer as to which is best yet.”

Overall pictureDetermining which feedstock to use in a given situationrequires looking at the overall picture. “For example, youneed to look at the social impact or the impact on jobs andthe health and safety aspects,” Osseweijer says. “And whatwe consider to be important here in the Netherlands may bedifferent to the people at the grassroots in Brazil. NGOs maycriticise the use of food crops for bioproducts, but Brazil hasa vast area of land available to grow products, and extensionof markets will help rural income. Food security is not reallyan issue here. Indeed, 40% of food produced in Brazil isexported. However, it does not mean that deforestation hasstopped, and we should therefore carefully analyse thesesocial impacts as well. For instance, most areas where sugarcane has been expanded show a higher humandevelopment index and higher education levels. For thelocals, it means a higher perceived level of well-being. Oneof our Brazilian students is looking at the actual andperceived impact of sugar cane at a grassroots level.”

Data gaps in the value chainJohn Posada Duque, also based in TU Delft, is working on themain data gaps in the value chain. The starting point involvesmore than 25 feedstocks from different locations in Brazil, fiveconversion technologies for jet biofuel production, and 10biobased products. "All of these options resulted in a largenumber of potential combinations for the value chain. So, wescreened these multiple options based on techno-economicand environmental criteria, but first we had to harmonise andvalidate the contradictory information obtained from theliterature,” he says. “We are now developing a framework todesign integral sustainable production scenarios with specialattention for social impacts assessment.”

Assessing the social, environmental and techno-economicimpact of jet biofuels and bioplastics

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Developing new methodologies and techniques is one aspect of driving forward sustainability in the biobasedeconomy. In the HIP project, however, the team has been looking at the techno-economic, environmental, andsocial consequences of producing jet biofuels and other renewables from a wide variety of feedstocks. Theultimate aim is to ensure the sustainable regional production of jet biofuels and bioplastics.

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FS5Microbial production of biofuels andbiorenewables

Upscaling the production of diesel and jet biofuels

Major steps forward have been made within Flagship 5’s project to develop an integrated bioreactor for theproduction of diesel and jet biofuels. The research team has developed technology to directly recover lipids used toproduce biofuels, a noteworthy result because both fermentation and separation take place in the same stage. In2015 the team realized proof of principle for the separation technology on fermentation broth and proof of conceptfor the reactor design. This has established the fundament to scale up the integrated process and equipment to aprototype of significant 100L scale. In 2016, this three metre bioreactor will be placed in the Bioprocess Pilot Facility(BPF), allowing larger volumes of diesel and jet biofuels to be produced. A successful 100L prototype run in 2017will be followed up by a pilot reactor of 8m3 at the BPF in 2018.

“In 2015 we focused on achieving two targets,” says KirstenSteinbusch, managing director of the BE-Basic startup DelftAdvanced Biorenewables (DAB). “An engineering post-docstudent from Delft University of Technology has worked onengineering and testing of the process and equipment forintegrated fermentation and separation of diesel and jetfuel-like biofuels. In addition, we focused on testing theseparation technology on real fermentation broth thatmicrobially produces diesel and jet biofuel-like compounds.And the ultimate aim was to integrate both parts and scaleup the integrated process and equipment to a 100Lbioreactor prototype. It was a terrific moment when werealised the separation technology really worked and whenthe plastic reactor prototype was given approval. It isfantastic that we now have the green light to build thereactor.”

Operational tests at the BPFThe team comprises three different groups working closelytogether: Delft University researchers within BE-Basic; DAB,the startup which is taking the promising technology to themarket and the BPF. “The pilot plant has been specificallydesigned to enable the transition from laboratory toindustrial scale,” says Peter Flippo, business developmentmanager at the BPF. “Over the past year DAB has carried out

operational tests with model emulsions containing twoprototypes from the DAB reactor. Being able to carry outfermentation and separation in one step is unique andreduces the scale of the process, so in the end you canproduce diesel and jet biofuels more cheaply. This isextremely important because of the pricing pressure in thebiofuels market.”

Full scale production in 2020The further upscaling steps that are planned are large scaleproduction at 1000L scale in a fermenter at the BPF. Thatreactor run will allow production of up to 1000L of fuel. “Byusing larger volumes, we can carry out more tests to validateour models,” says Steinbusch. “In addition, we can use theprinciple of fermentation separation to carry out tests usingother organisms. At DAB, we will also do this on behalf ofthird parties, who will also be able to upscale using a reactorinstalled at the BPF.” The development is good news for theproduction of diesel and jet biofuel in general. “Once theupscaling phase has been completed at the BPF by validationof a second scale-up step to 8000L by 2018, by 2019 itshould be possible to build a factory,” says Steinbusch.“Industry itself will have to actually set up a full-scale plant,but the fuel could be produced on a commercial scale from2021.”

Bioconstruction materials The last stage before a major market launch

The self-healing concrete developed by Henk Jonkers of Delft University of Technology has been creating a buzz foryears, and in 2015, Jonkers was nominated for a European Inventor Award by the European Patent Office.Following the development of the basic idea, Jonkers has been working with Corbion, to develop a fully-fledgedproduct. Companies from all over the world are interested, but a commercial launch remains a challenge.

“We made a major advance from the lab phase to actualprojection in 2015,” says Henk Jonkers. “We can nowproduce several thousand kilos of self-healing agent,bacterial spores, nutrients and lactate/polylactic acid-basedsubstrate, which heal the concrete from within – that’senough for 20,000 kilos of concrete. The only problem is wecurrently need two external companies to produce a ready-to-use product and that is forcing the price up to some €80extra per cubic metre of concrete. This is too expensive forthe construction industry.”

Reducing the priceNothing more can be done to cut the price of the healingagent. “From a scientific perspective, it has been developed.We have the right bacteria, the right product compositionand the right processes,” says Jonkers. “The price is due tothe raw materials and the fact we need outsiders in theproduction process.” The solution would appear to bestraightforward: start their own healing agent productionline. “That is a serious option,” Jonkers says. “It could takethe price down to €30 a cubic metre of concrete and that isacceptable to the private sector. But it would requireinvestment of €4M. There are potential investors, but theyquite rightly want confirmed orders before they start.”

Show it worksProof that the self-healing agent works is one of the majorvalue adds that Jonkers’ research group has. “Concreteoften gets damaged warming up after frost. We’ve beenable to speed this process up in the lab to show the positiveeffects. But commercial companies really want to see the

process in reality. So we are doing as many practical tests aswe can. For example, the Limburg Water Board has a wastewater processing tank which is partly made of self-healingconcrete. The tank looks fine at the moment, but of courseconcrete rot takes a few years to materialise. So we don’tknow when we will see that the self-healing section is inbetter condition. The water board has already said that assoon as the process has been proved, they will use thehealing agent in all their projects. Then you start to makemajor advances.”

More speed using repair mortarThe healing agent can also be added to the mortar used torepair current damage, and Jonkers is running practical teststo prove this to potential buyers as well. One of theseinvolves a project in Groningen for construction group BAM.“You can show the way the healing agent works in therepair mortar more quickly, and companies are prepared topay more for it. We expect that the repair mortar will beaccepted more readily by the market and that buyers willthen be willing to put the healing agent in new concrete.That will take setting up our own production line a stepcloser.”

The self-healing concrete is based on bacteria which produce calcite when fed and activated. The bacteria and its foodstuff (the healing agent) are added to the concrete in tiny capsules that only open when the concrete hashardened and cracked. The capsules are made frombioplastic developed by Corbion.

FS4The Flagship Bioconstructionmaterials focuses on the developmentand testing of microorganism-basedsolutions to create smart(bio)construction materials which areself-healing.

The Flagship Microbial production ofbiofuels and biorenewables focuses onthe development and optimisation ofmicrobial-based conversion processtechnologies for the conversion ofbiomass into biofuels and otherbiorenewables.

Synthetic biologyThe Flagship Synthetic biology develops tools and techniques for the improvement of micro-organisms. The design and optimisation of novel pathways to desired products are complemented by unique cellmembrane engineering aiming for efficient product export and improved robustness of the productionorganisms.

New form of lifeThe idea behind the Flagship’s work is to make E.colibacteria more robust. Every cell’s membrane is composedof lipids and the lipids, in Archaea are thought to beresponsible, at least in part, for the organism’s ability tolive in such extreme conditions. “So if we can introducethe Archaea ether lipids into the E.coli membrane, weshould be able to improve the bacteria’s robustness,” saysCaforio. Once this has been achieved, the team will havedeveloped an E.coli bacteria which should have a highertolerance to toxic products, organic solvents andbyproducts without loss of productivity. Such anorganism with a mixed membrane will be a new form oflife. “The existence of a last universal common ancestor(LUCA) with a mixed membrane is one of the hypothesesat the basis of the differentiation between Archaea andbacteria,” says Caforio. “The development of a viablebacterial cell with a mixed membrane will help usunderstand the molecular basis of evolution.”

Methodology to engineer lipid speciesIn 2015, the team succeeded in identifying all the keyenzymes involved in the ether lipid pathway and thepathway, was introduced into E.coli via metabolicengineering. The development can be seen as a generalmethodology to engineer lipid species that can beapplied to other micro-organisms which have industrialrelevance. In other words, it has opened the door tofurther improving the process and proving a directcorrelation between the introduction of the ether lipidsand the robustness of the E.coli membrane. “We could

detect the presence of the archaeal lipids in the E.coli butnot in sufficient amounts,” Caforio says. “We are nowimproving the strain and optimising the processes to tryto produce higher production of lipids and reallydetermine if the process has improved the robustness ofthe cells. So far, our efforts are paying off.”

Actual applications“In previous years the focus was on the production ofether lipids in the E.coli membrane to try to affect theproperties of the bacteria. Now the robustness tests arebeing performed on the evolved bacteria strain and,depending on the results, will be used to develop actualapplications,” says Caforio. Eventually, the techniquecould be applied to other industrially relevantmicroorganisms to increase high temperature or pressurestability. The robustness of the constructed strain couldalso support developments in lipid-based drug deliverysystems. For example, if the carrier is made more resistantto the acidic environment of the gastro intestinal tract,drugs could be delivered to the targets without beingcompromised.

Helping cells to survive in more extreme conditions

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FS6

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Can cell membranes be engineered to become more robust and survive in more extreme conditions, such ascold and heat, and can they be made tolerant to organic solvents or byproducts during industrial processes?“Archaea are single cell organisms that can withstand really difficult environments such as hot springs orextremely cold habitats,” says Groningen University researcher Antonella Caforio, who has been working on oneof the projects in this Flagship for nearly four years together with a team in Wageningen. “If we can introducethis ability to other organisms such as bacteria, we may be able to make them withstand stressful conditionsand, in turn, increase cell productivity during industrial processes.”

High-throughput experimentation andmetagenomic miningThe Flagship High-throughput experimentation and metagenomic mining develops and applies high-throughput approaches and tools to explore and mine the metagenome, genetic material that comesdirectly from our natural ecosystem. In addition, this flagship aims to engineer and screen enzymes andother products for improved properties.

Competition between bacteria“When they are in pure culture, many bacteria do nothingbut when they notice another strain, they start tocompete and for this often produce bioactivecompounds” says Flagship manager professor Hans vanVeen of the Netherlands Institute for Ecology, NIOO-KNAW, which is based in Wageningen. “We’ve discoveredthat bacteria turn different groups of genes on and off toproduce various substances, depending on theneighbouring bacterial species. High-throughputscreening is helping researcher Olaf Tyc to identifyinteresting combinations of bacterial species capable ofproducing novel bioactives.”

The project is part of Flagship 7 and aims to identifypromising new compounds. “We have also looked atrotting wood because we expected to find usefulantioxidants which pose less of a risk to eukaryotic cellsand therefore ultimately to humans,” says Van Veen. “Thetheory did not entirely work out, but we did find acompletely new family of bacteria which are closelyrelated to the commonly used antibiotic-producingStreptomyces. We’ve managed to map the genomes ofseveral species of this family so that in the future we canscreen for the relevant characteristics more quickly andaccurately. That research is now being continued in closecollaboration with Leiden University.”

Towards the marketThe search for new compounds within Flagship 7 will endmid-2017. "Finding the substance is only the first step,”says Van Veen. “After that, we face the challenge ofobtaining the pure compound from the bacteria andbuilding up a file of useful information for the market.”Van Veen sees his mission as to bridge the gap betweenscience and the market. “Scientists no longer sit in ivory

towers,” he says. “Everyone who works here is aware ofthe need for practical applications. Of course, it would begreat if my research led to the launch of a new antibioticor if the novel antioxidants were used in cosmeticproducts. But here at this institute, whose core activity isfundamental ecological research, we cannot carry outthousands of tests if they don’t contribute to improvingour fundamental knowledge and to our main products:our publications and theses. And the market needs thosetests before they will invest. We are trying to close thatgap with start-ups such as MicroLife Solutions (MLS), andwe are increasingly talking to industry at an earlier stagein the research process. This is how we try to encourageresearchers to become more market-oriented and tointerest industry in investing in highly promising research.For example, Dick Janssen’s research on modellingenzymes in another Flagship 7 cluster involves intensivecontact with industry and has generated patentapplications.”

Towards the futureInteresting discoveries continue to be made, even thoughthe project is nearing its end. “We’ve found Acidobacteria which produce new forms of PHA and EPS, bothof which are bioplastic components and non-toxic inthese forms. They offer real commercial opportunities,”says Van Veen. Such discoveries, illustrate the added valueprovided by BE-Basic. “The charm of a big project like thisis that you can carry out broad-based research. BE-Basichas given metagenomic mining an enormous boost andshown which research angles and methods have the bestchance of success. There is an incredible amount todiscover in nature, and high-throughput genomic researchmakes those opportunities more visible. I’m convinced itwill lead to the delivery of many more commerciallyinteresting concepts and products.”

‘As there is an immense societal demand for new bioactive compounds such as antibiotics and novel enzymes to be used in biobased production processes, it is fantastic to identify a novel microbial compound which becomes a concrete product.’

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The hunt for new and improved enzymes and bioactive compounds continues to deliver surprises. Take, forexample, the discovery that two different bacteria growing in association produce a compound active againstfungi and pathogenic bacteria, which is not formed when the organisms are grown separately. Such compoundscould be used as antibiotics. High-throughput technology makes it possible to rapidly test different combinationsof bacteria.

FS7

Environmental impact of chemicals, biobased molecules and processesThe Flagship Environmental impact of chemicals, biobased molecules and processes develops novel andefficient biobased monitoring tools for the evaluation and improvement of chemical safety in the biobasedeconomy. Research is focused on the environmental and human safety issues arising from the transition toand implementation of a biobased economy, as compared to existing industrial activities.

Professor Wim van der Putten of the Netherlands Institutefor Ecology NIOO-KNAW, Flagship manager professorHauke Smidt of Wageningen University, and doctor Bartvan der Burg from BDS have been looking back togetherat the results booked by Flagship 8 in 2015. “After all,”says Van der Putten, project leader within FS8 andmanager of FS3, “it is the variation of expertise withinBE-Basic and the willingness to cooperate acrossdisciplines and flagships that are key to our success. Forexample, FS3 is able to use techniques and tools thathave been developed in FS8 and that are now beingtested and integrated into decision support systems.”“It has also helped in the way we think,” adds Smidt.“The starting point for this flagship was ‘end of pipe’ – towarn about and clean up dangerous substances. Now wehave a different perspective. Perhaps we could createsomething new and sustainable by seeing waste as aresource instead.”

Variety and precision in farmingTake recent research by Van der Putten and colleagues,for example, which establishes that willow can improvesoil. “By rotating between maize and willow you actuallyimprove impoverished soil and produce raw materials tomake bioethanol at the same time,” says Van der Burg.“The knowledge we have now enables us to look two tothree steps ahead,” says Van der Putten. “That isnecessary in order to design an entire production chain,including bringing value to the waste products.” Scientific advances have also been made in remotesensing, which is key, for example, to developing adecision support system for farmers. Remote sensingallows farmers to decide how to manage crop health byanticipating future problems. “We have established proofof principle that a different microbial make-up in the soilcan be detected by hyperspectral imaging of the plant

itself, by measuring wavelengths that cannot be seen byus humans,” says Van der Putten. “This offers greatperspectives for precision farming, for example byhelping farmers to decide where on their land they canmake use of which type of waste.” It will still take sometime before this becomes a common practice. “A lot ofresearch still has to be done, and a market launch takestime,” says Smidt. “But it is an important developmentfor the decision support tool.” “These discoveries,” says Van der Putten, “could have agreat impact on the sustainability of human society.”

ScreeningIn the meantime, BDS has won its spurs with the CALUXscreening panels that highlight toxic substances byilluminating cells. “We can rapidly identify biobasedcompounds with low toxicity,” says Van der Burg. "Wehave recently shown that biobased FDCA (furan-2,5-dicarboxylic acid) is an interesting new 'green' alternativeto plastic building blocks as it is devoid of endocrine andother activities often associated with these molecules. Inaddition, we are trying to adapt the technique toincreasingly complex blends. In the end, you want to beable to test biobased waste streams quickly, and bydefinition these streams contain biological activity. Thescreening panel should make it possible to separate thebeneficial biological activities from the harmful. Behindthe scenes we are making great steps forward in this.”The team’s vision for screening tools has also changed.“The tools focus on the point at which a substancebecomes toxic,” says Van der Burg. “But before that pointis reached, the substance might be beneficial in smallerquantities.” Smidt adds, “Our perspective has shiftedfrom warning about dangers to identifying economicallyvaluable molecules in biomass. This is essential to makethe biobased economy a real option.”

From monitoring chemical safety to creating added value

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Flagship 8 focuses on developing tools and methods to measure whether biobased building blocks are safe formankind and nature and how to map the risks associated with soil exhaustion. Now more results are beingevaluated, this risk analysis has become the starting point for adding value across the chain.

FS8

“The conclusions of the peer reviewed report can be calledspectacular,” says Osseweijer. “It concludes that landavailability is not a limiting factor and that bioenergy cancontribute to sustainable energy and food supplies evenwith increasing food demands, the preservation of forests,protected lands, and rising urbanisation.” The report waslaunched in São Paulo, Brussels and at the World Bank inWashington. “Such extensive communication is crucial,and it is good to see so many people who are involved inbioenergy innovation use the report. A policy paper basedon it was presented during the European launch at theGreen Energy Week in Brussels.”

New tools to boost cooperationThe SCOPE report also reviews studies on sustainabilityimpacts and modelling, including those carried out withinthe confines of BE-Basic. “Our programme is crucial tohelp innovation succeed,” says Osseweijer. “And with theresults now being presented, we see increasing attentionfor the subject, from industry as well. This year a numberof sustainability studies led by the Copernicus Institutewere finalised, providing models for sustainable land usefor biobased production. The programme takes a verybroad approach and looks at the influence of land usechanges, the socio-economic aspects and smartmonitoring tools. Such tools should help stakeholdersthroughout the value chain to decide on the bestpractices for crop management, transport, and biobased

production. This is unique and necessary for thetransition from a fossil-based to a biobased economybecause society demands this be sustainable. Also,situations are radically different in a biobased economy.For example, the chemicals industry needs to link withfarmers for feedstocks – and that requires different toolsfor cooperation and agreements to ensure a reliablesupply of feedstocks.”

Biorefineries, the harmonisation of certification andeducationMore is needed to support this transition, includingensuring that the new insights land on the right desksand that future engineers and entrepreneurs are welleducated and trained. For example, BE-Basic hasprovided input for the ‘redefinery’ – a blueprint forbuilding a biorefinery in the Netherlands whichincorporates the impact of socio-economic factors. Theharmonisation of European rules for certifying solidbiomass is another aspect which BE-Basic contributed to.This is key to organisations which issue sustainabilitycertificates. Furthermore, extensive efforts have beenmade to boost both professionalisation and digitalisationin education. “We want to learn from our success withthe MOOC and digitalise educational elements that canbe used in different settings, and for large audiences,such as preparatory material for an advanced Master orPhD course,” says Osseweijer. “Our second MOOC

‘Availability of land is not a limiting factor: will this finally bring the biobased economy closer to reality?’

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In 2015, BE-Basic booked its best ever results in the field of embedding the biobased economy in society at large.It was the year the SCOPE report on Bioenergy and Sustainability, which offers a positive perspective on therealisation of a global bioeconomy, was published. The 800-page report by the Scientific Committee on Problemsof the Environment reviews scientific evidence on the benefits of sustainable energy to society. “The report is alreadybeing called the ‘Bible of sustainable bioenergy’,” says Patricia Osseweijer, chairwoman of the Economy, Policy andSustainability (EBD) programme. “The peer-reviewed report is a collective effort with contributions from 137researchers at 82 institutions in 24 countries, and we are proud that BE-Basic experts contributed to nine of the 21chapters. The impact is immense because the results give countries and companies a license to move forward.”

FS9FS11Societal embedding of a biobased economyThe Flagship Societal embedding of a biobased economy optimises the societal embedding of the productsand processes developed by BE-Basic. The Flagship focuses on the identification of socio-economic aspects andsustainability issues and the development of adequate systems to monitor and model them. The Flagshipdevelops effective and efficient education, communication and societal valorisation programmes.

Flagship 9 and Flagship 11 both focus on societal aspects of importance for the transition to a biobased economy.

Corbey CommitteeThe Corbey Committee called on the Dutch government tomake developing a biobased economy a much higherpriority with the publication of its vision document ‘Naareen duurzame bio-economie’ (‘Towards a sustainablebioeconomy’) in October 2015. The document points outthe advantages of a bioeconomy in which biomass is aresource: fewer CO2-emissions, less dependency on fossilfuels, and more employment opportunities. It explains thatthe biobased economy would be lucrative for theNetherlands, but that it requires ambitious integralpolicymaking and a stronger government role to drive itforward. The results of two BE-Basic research projects onthe availability of national biomass and the import ofbiomass into the Netherlands were included in the report.

International Energy AgencyBE-Basic supports the Dutch membership of theInternational Energy Agency (IEA). IEA Bioenergyʼs strategicplan envisions biomass as making a substantivecontribution to future global energy demands byaccelerating the production and use of environmentallysound, socially accepted and cost-competitive bioenergyon a sustainable basis. This would provide increasedsecurity of supply and reduce greenhouse gas emissionsfrom energy use. Dutch industry and research institutes areinvolved via an implementing agreement, financed by theTKI-BBE.

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BE-Basic’s Macroeconomic outlook of sustainable energyand biorenewables innovations (MEV II) was completed in2015. The extensive study offers guidance for Dutch energyand bioeconomy strategy. Research leader Hans van Meijlfrom LEI Wageningen UR discusses the main findings.

1. What is the Macroeconomic Outlook II?The study looks at the impact of a Dutch biobased economyin which biomass is used on a major scale, both at a systemand a macroeconomic level. System-level impacts includerenewable energy deployment, fossil fuel reduction, andgreenhouse gas (GHG) mitigation. Impacts at themacroeconomic level include gross domestic product (GDP),value added, employment and trade balance.

2. What are the most important findings?The study shows that the biobased economy can make apositive contribution to the Dutch economy in general andwill help cut greenhouse gas emissions as well as cuttingthe cost of achieving that reduction. The macroeconomicimpact depends on fast technological change, high fossilenergy prices, a worldwide trading system and low biomassprices. A biobased economy which is driven by fasttechnological change and cheap, easily available biomass,will cut the cost of reducing CO2-emissions and will makebiobased options more attractive. But the bioeconomy willnot become a reality while oil prices are at their currentlevel. Intervention in the form of a tax on CO2 or changes inR&D strategies would have an impact on this.

3. What are the main factors influencing the transitionto a biobased economy?There are a number of issues which need to be taken intoaccount in achieving the above-mentioned results andswitching to a biobased economy. Major technologicaladvances, high fossil energy prices and the low worldwideprices for biomass are essential elements for a successfultransition. At the same time, low fossil fuel prices dampenthe macroeconomic advantages.

4. What more can be done to drive thesedevelopments forward?The study shows that a strategy of stimulation – such asCO2 taxes or R&D policy – is key to realising themacroeconomic benefits and a reduction in CO2-emissions.For example, the Netherlands supports electricityproduction using wind and thermal energy and that hasmade a considerable contribution to meeting targets onrenewable energy sources. In addition, technologicaldevelopments are needed within biobased sectors to makebioenergy and biochemicals cost competitive. Biorefineriescan play a crucial role here. A high tax on CO2-emissionscan also be an effective policy instrument. Low feedstockprices and good access to international sources of biomassare also essential. So stable international biomass marketswill need to be established as well.

5. How do the BE-Basic findings impact the realisationof a biobased economy?BE-Basic stimulates the development of fundamental andapplied technological knowledge to make biobasedtechnologies a competitive reality. This is essential in thetransition to a biobased economy. In addition, BE-Basicfocuses on driving forward social acceptance of thechanges. This is crucial, as the debate on GMOs withinEurope has shown.

Five questions about the Macroeconomic Outlook II

PartnersThe research was carried out by LEI Wageningen UR andthe Copernicus Institute at the University of Utrecht.Project partners Essent, DSM and Corbion alsocontributed to the economic and technical aspects ofthe report. The public version of the impact study andthe full scientific report can be downloaded for freefrom: http://www.be-basic.org/downloads.html.

FS9FS11

‘Industrial Biotechnology’ was followed by 10,000 students.And in Brazil, many courses have been developed which weoffer to a wider public using different forums.”

Focus on communicationIn the meantime, a broad discussion about the biobasedeconomy is under way in society at large, helped bypublications such as the SCOPE report. BE-Basic has beeninstrumental in driving the debate forward by, for example,publishing a weighty reaction to the Dutch Economic AffairsMinistry’s new energy policy. “There are many newinsights,” says Osseweijer. “We know more about thecritical points in the chain and which technologies arefeasible. And we’ve debunked several myths. One of themost stand-out facts is that we do have enough land to feedthe growing population and that the global food versus fueldiscussion is history in terms of facts. For example, it hasbeen established that the use of land to grow biomass forfuel production actually can improve local food productionand quality. Growing raw materials for bioenergy inducessustainable agriculture and that means people get betteraccess to food. So there are improvements across the entirechain. We need to communicate these insights to a muchbroader public to give the biobased economy a real chanceof success. So in 2016, we are going to enlarge our focus oncommunication.”

Developing policyMaking the biobased economy a reality also demands achange in official policies. Within BE-Basic, Flagship 11 hasdeveloped various models and tools to calculate variablessuch as emissions and to predict the economic impact ofdifferent scenarios. The Macroeconomic Outlook waspublished in 2015. BE-Basic also contributed to the CorbeyCommittee report on the availability of biomass. “Thecombined projects of our Flagships are accumulating abetter understanding of how novel technology can land insociety in a sustainable way and how we could helpaccelerate positive impacts,” concludes Osseweijer.

EBD Programme: Economy, Policy andSustainability Flagship 11 EBD Programme: Economy, Policy and Sustainability focuses on providing insight into the societalpreconditions for the economic and sustainable introduction of bio-renewable chemicals, materials, fuel andenergy production. It provides policy recommendations on the likely macro-economic impact of the large-scaleuse of biomass in the Netherlands (including imports) and aspects of sustainability and contributes to(inter)national committees.

Flagship 9 and Flagship 11 both focus on societal aspects of importance for the transition to a biobased economy.

The novel strain improvement project is very much drivenby industry. “Lactic acid bacteria determine the taste,texture and preservation of dairy products,” Kuipers says.“The food industry is interested in other flavourcompounds and improving anti-bacterial activity to, forexample, increase the shelf life of a product. The foodindustry would be extremely pleased if we can adapttransfer processes that occur naturally.” Four PhDresearch projects are now underway which focus on threeof the bacteria which occur in cheese, yoghurt or otherfood: Lactococcus lactis, Streptococcus thermophiles andBacillus subtilis.

Mapping the transfer of genesThe researchers have looked into three ways by whichgenes are transferred horizontally in nature, and Kuipersand his team are now developing the technologicalmethodology to do this in the laboratory. “First of all,bacteria can transfer DNA material via a process knownas conjugation. Then there are bacteria which take upDNA out of the environment in a process which we callcompetence. This happens naturally, for example, duringdecomposition in soil. Thirdly, there is phagetransduction, a natural process in which foreign DNA isintroduced into a cell. Phages are bacterial viruses thatinfect bacteria and take over part of the DNA – this isalso something which happens in soil. To carry out thisresearch, we have access to a large collection of phagescapable of transferring DNA. This is a process we canmimic in the lab and stimulate.”

Selecting characteristicsFurther development is needed to ensure the technologycan be used by industry. “Transferring DNA is not theproblem,” says Kuipers. “The challenge now lies indeveloping ways to select the characteristics which youwant to obtain. We want to develop naturalmethodologies, so we want to carry out this screeningprocess in a natural way as well. These characteristics canbe captured, for example, by screening for phenotypicalmarkers. We could for instance make selections based onpolysaccharides, which can be easily identified becausethey create slimy colonies. But we would rather hone indirectly on DNA, which is why we are researching the useof specific probes."

Possible industrial and environmental applicationsDevelopments in this field are being closely followed bythe Flagship’s industrial partners. “We only take ourresults to the outside world once they have been clearedby industry because they could create major shifts in themarket. The technology we are developing could also beused for other applications. For example, certain bacteriaand fungi could contribute to clearing up pollution. It isquite amazing what natural methodology makespossible.”

Seeking a natural way to improve lactic acid bacteria

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Is it possible to improve strains of lactic acid bacteria using methods that also occur in nature? This question isat the heart of the novel strain improvement project within Flagship 10, led by Oscar Kuipers at the University ofGroningen in close cooperation with several industrial partners. “The project has been running for under twoyears,” says Kuipers. “We have looked at three natural methods for transferring lactic acid bacteria DNA andhave developed new methods to achieve strain improvement in the laboratory. In time, this will help industry todevelop new flavours and boost the shelf life of dairy products.”

Genomics for industrial fermentationThe Flagship Genomics for industrial fermentation aims to deliver improved fermentation processes forbioconversions in the food industry, focussing on the selection of natural strains or strains resulting fromdirected evolution or (non)-genetic modification.

FS10

Iso-Butanol Platform Rotterdam (IBPR)

The Flagship Iso-Butanol Platform Rotterdam (IBPR) explores a biobased value chain for producingcommodity chemicals and fuels from woody biomass using an improved Organosolv pretreatment and isbased in the Rotterdam region.

Technical feasibility biorefinery demonstrated

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Flagship 12’s main project focuses on the cascading conversion of lignocellulosic feedstock into isobutanol andthe further valorisation of its derivatives as well as developing high-value applications of the lignin fraction. In2015, the team was able to demonstrate the technical feasibility of a biorefinery in the Rotterdam area based onthe platform molecule isobutanol. The project encompassed the entire value chain from lignocellulosic biomassup to and including the high-value end-products.

FS12Researchers Jaap van Hal, innovation manager at ECN,and Ana López-Contreras, senior scientist in foodbiobased research at Wageningen UR, say they arepleased with the results so far. “The results show theimportance of isobutanol as a platform molecule,”confirms Van Hal. “In this project all the individualprocess steps of the entire value chain of the biorefineryhave been proven on lab scale. And the economics of thefull-scale biorefinery were estimated in detail based on adedicated process design.”

Three main fractionsDuring the conversion process, the biomass isfractionated into three main fractions: cellulose pulp,hemicellulose, and lignin. Isobutanol is produced byfermenting the fractions which contain carbohydrates.The end products studied in the project are GTBE,isobutyl acetate and isobutanol-acetone condensate, aswell as the purified lignin that results from the ECN-patented, ketone-based Organosolv pre-treatmentprocess.

The best case plant design shows a feasible pay-out timeof six years. The life cycle assessment shows theadvantages of the biobased isobutanol platform incomparison to the petrochemical isobutanol platform.The ‘non-renewable energy use’ is approximately 45%lower when compared to its petrochemical counterpart.The GHG emissions are up to 25% lower for thebiorefinery when compared to their equivalent fossilsystems. For fresh water depletion, savings can even beup to 58% over their petrochemical counterparts.

Room for further optimisation"The project was successful,” says López-Contreras.“We've got a good hydrolysis and fermentation processfor isobutanol. The fermentation can be optimised stillfurther for the use of highly concentrated sugarsolutions, because of the toxic components for the micro-organisms in these solutions."

The economic viability hinges on developing high-valueapplications of lignin. This lignin fraction is currentlyburned in most pretreatment processes, but scientists arealso looking at whether it is possible to give the high-purity lignin of this process added value so that it can beused, for example, in coatings or bunker oil. The nextstep is the development of a pilot plant based on abusiness plan for a world-scale plant in the port ofRotterdam.

The Flagship 12 consortium members are: BE-BasicFoundation, AVR, Corbion, Deltalinqs, ECN, GEVO,Sweco (formerly Grontmij), Port of Rotterdam, Procede,Delft University of Technology, Utrecht University, Foodand Biobased Products Wageningen UR, BDS andZirk©Technology. In addition, the IPBR project wassupported by EFRO (Europees Fonds voor RegionaleOntwikkeling).

Investing in your futureThe IBPR project is partly financed by theEuropean Development Fund of theEuropean Union

BE-Basic Foundation Annual Report

International alliances: seizing opportunities

VietnamThe research into intelligent mining in Vietnam has broughtinto focus various enzymes capable of breaking downcellulose and lignin. These enzymes contribute to the pre-treatment of biomass for biofuel, a process which isnecessary to make the production process faster and moreprofitable. “We have demonstrated proof of principle in thelaboratory, a breakthrough I’m very happy about,” saysBram Brouwer, general director at MicroLife Solutions andmanaging director at BE-Basic. The enzymes were found inhot springs and mud pools in a remote, almost uninhabitedarea of Vietnam. “Nature has had its way there undisturbedfor years, allowing the rotting leaves of the trees to create aspecial microbial environment in the mud pools. We thinkwe will be able to find similar enzymes in the hot springs,with the additional advantage that these enzymes are activeat temperatures of 60° to 90° Celsius. This means they willbe able to withstand comparable temperatures in the pre-treatment reactors. We are also looking at enzyme geneticswhich can help determine evidence of clusters of propertiesin new microorganisms in order to speed up discovery.”

Sustainable researchThe breakthrough has been welcomed both by Vietnam andthe Netherlands. “Now we need to work towards a pilotand a production environment, but we need to act carefullyand respect nature,” says Brouwer. “You can’t just rip outlots of plants, or sample many mud pools formicroorganisms because before you know it, you will havedestroyed a unique environment. We are talking aboutsetting up a system for growing plants and microorganismson location in a national park which we think could beoperative by the end of 2016 or in 2017. BE-Basic ratessustainability highly, a fact much appreciated by ourVietnamese partners. We are even held up as examples toneighbouring countries.”

EuropeCooperation between theNetherlands (BE-Basic), Flanders(FISCH) and North Rhine-Westphalia (CLIB2021) in theBioInnovation Growth Cluster (BIG-C) has become more concretein 2015. BE-Basic Managing Director Luuk van der Wielen isclosely involved in BIG-C. “There is a lot of industrial activityin this region, much of it dependent on fossil fuels,” he says.“The better you can tap into the existing infrastructure, theeasier and more successful the transition to a biobased wayof working will be. Is it possible to extract feedstock fromfossils, or use carbon as a raw material for new processes?These are the kinds of questions we need to find answers to,and technological and stakeholder analyses are alreadybeing made to determine the role of carbon in this area aswell as the extraction of aromates from wood biomass.” Theproject is very much an international endeavour. “Europeancooperation is essential,” says Van der Wielen. “Many of theBIG-C projects are developed along European Union researchlines. And that research base is being broadenedcontinuously. Recently, we have entered into a partnershipwith Toulouse White Biotech so we can share knowledgeand facilities. We have also agreed with CLIB to initiateinnovative partnerships with Brazilian organisations in orderto speed up developments in Europe.”

BrazilOpportunities for the development and implementation ofbiobased solutions continue to present themselves in Brazil,even if the economy is not growing as fast as it has been.The 2015 public-private partnership Agropolo, modelled ona French initiative, is just one of the many innovativeventures initiated here. Agropolo is not only about sciencebut also looks at the social implications of the agro-industry.“The Brazilians have come to the conclusion that theindustry needs reforming. At the moment it is all about endproducts, such as citrus juice or meat for consumption. Nowthey want to look into ways of making waste flowsprofitable as well,” Van der Wielen says. The first phase ofthe Agropolo project consists partly of workshops explainingwhat might be done. BE-Basic has ample experience inpublic-private partnerships in this area and is closely involvedin this part of the project. “We help develop ideas from stepone. The process is similar to that involved in theMacroeconomic Outlook (see page 29) in the Netherlands. Ifyou want a biobased economy with a certain GDP, a certainlevel of employment and certain emission standard, how doyou go about it? What links are missing in the value chain?Opportunities become feasible if you back them up withfigures, and that is what is happening, for the first time, inthe region of São Paulo. Eventually, it will happen in thewhole of Brazil.” Van der Wielen thinks Dutch industry couldbenefit from the developments in Brazil. “Together with theNetherlands Enterprise Agency (RVO.nl) we are looking intothe instruments needed to help companies wanting to workthere, such as subsidies and practical support.” BE-Basic hashad an office in Brazil for a number of years, and that is agreat advantage, too. “We have our own networks as wellas the ones provided by the Dutch government,” he says.

Dual degreesOne of the key objects of BE-Basic Brazil is to put in place adual degree structure which will see people take PhDdegrees both in Brazil and the Netherlands. The hard work isbeginning to bear fruit. “Many people are very interestedindeed,” says Van der Wielen. “High level education is acritical priority for the future of Brazil.” This was underlinedby over 30 Brazilian and Dutch academic leaders during arecent visit to Brazil by Dutch Education Minister JetBussemaker. “The minister was very enthusiastic about theprogramme,” says Van der Wielen. “We have now agreed toextend the dual degree programme from 20 places in 2020to 100 by 2025. At this moment only TU Delft and theUniversity of Campinas UNICAMP offer a dual degree routebut there is room for expansion and others will follow. Theadded value of a dual degree construction is beingrecognised more widely. It would be nice if the University ofWageningen would agree to come in as well. Most biomassis found outside of the Netherlands, that we know. So it isup to us to go out there and take advantage of that fact todevelop a bright, biobased future.”

34

A breakthrough in enzyme research in Vietnam, concrete feasibility studies in Brazil, broader cooperationwithin BIG-C, and over 25 dual degree students working on projects or preparing to do so: 2015 has been agood year for international alliances.

35

MOOCIn 2015, BE-Basic’s Massive Open OnlineCourse (MOOC) on ‘Industrial Biotechnology’(formerly ‘Technology for biobased products’)was run for the second time through the EdXplatform. In the MOOC, biotechnology,bioprocess design and sustainabilityassessment are integrated into 66 videolectures by course leaders from TU Delft andguest lectures from industry and UNICAMP.The second edition attracted about 10,000participants – in line with the first edition in2014. In total, 100 of the participants receivedan ID-verified certificate and 225 received a(free) honorary certificate.

Bioprocess Pilot Facility openedThe Bioprocess Pilot Facility (BPF) was officiallyopened in March 2015. The BPF is a uniquemulti-purpose facility for bioprocesses.Universities, companies and institutions fromall over the world can use this open accessfacility to investigate how productionprocesses can be scaled up. The BPF is locatedat the Biotech Campus Delft and is allied withBE-Basic Foundation (see also Flagship 5). Inthe early plans, the pilot plant was part of theBE-Basic programme, but has since beendeveloped as a separate entity by itsshareholders Corbion, TU Delft and DSM.Several BE-Basic projects will test their newtechnologies at the BPF.

3736

Activities

BE-Basic Annual MeetingThis year the BE-Basic Foundation celebrated its 10th anniversary. Inrecognition of this, a special edition of the public Annual Reportwith a video highlighting biobased product, process and companydevelopment over the last 10 years was launched at the annualgeneral meeting. The event at the Leeuwenhorst conference centrein Noordwijkerhout, was attended by some 250 guests. It focusedon the consortium’s recent developments and on the next stepsneeded to implement biobased technology into society.

Guest speaker Erick Fernandes (World Bank) reflected on the rapidlygrowing world population and the increasing demand for meat,(bio)fuels, food and energy. Sean Simpson (LanzaTech) and Niels vanStralen (ChainCraft) focussed their presentations on how to bridgescientific knowledge and concrete entrepreneurship.

This year’s winners of the Poster Pitch and Poster Award were Yourivan Nuland and Willem Dijkman, respectively.

Holland Biotech Pavilion in MontrealBE-Basic again coordinated the Holland Biotech Pavilion at the BIO World Congress on Industrial Biotechnology,which took place in Montreal between July 19 and 22. It is the third time in a row that BE-Basic has overseen thecoordination. The pavilion included 12 Dutch organisations and companies with a focus on biotechnology,biochemistry, energy, feedstock production, transport and logistics. The Holland Biotech Pavilion is financiallysupported by the Topsector Chemie via the Strategic Conferences Program coordinated by RVO.nl.

With more than 1,200 visitors from some 725 organisations from around the world, the congress was well attendedand offered an excellent opportunity to explore mutual interests for international partnerships through one-on-onepartnering meetings and network receptions. Furthermore, BE-Basic organised a partnering workshop to giveCanadian and Dutch organisations an opportunity to pitch their business. The bike raffle, presented by Rochus Pronk,Deputy Head of Mission at the Dutch Embassy in Canada, was sponsored by the Rabobank and proved a greatsuccess.

After the BIO World Congress, a delegation from the Netherlands travelled to Sarnia in the province of Ontario for aworkshop and a company tour, organised by Bioindustrial Innovation Canada.

BE-BIC

The BE-Basic Innovation Centre (BE-BIC) stimula

tes and supports all

the innovation activities taking place within the

BE-Basic research

programme. In 2015 BE-BIC organised the fifth

and final Ideation

Workshop for postdocs and PhD candidates to

raise awareness of

how technological know-how can be turned int

o business. In

September 2015, the first Action Lab took plac

e: five teams of

researchers developed a business plan and pres

ented the result to

the management team. The winning team from

Wageningen

University presented a business case for their ne

w technology to

discover plant enzymes that can be used to pro

duce natural flavours

by fermentation. Their prize was €20,000 to fu

rther develop their

ideas within their spin-off Radix Botanicals. Two

other teams,

MicroLife Solutions and DAB (Delft Advanced B

iorenewables), each

won an incentive prize of €10,000.

One participant, Bart Pieterse (Green Waste Su

pport) explains: “The

Action Lab was very valuable for me. Without t

he right focus, you

can easily waste a year of research.”

38

i

60

50

40

30

20

10

0

2004

2005

2006

2007

2008

2009

2010

2011

2012

2013

2014

2015

2016

2017

2018

2019

= Forecast

= EU & National projects 2011-2015

= EFRO-IBPR

= Brazil activities

= Vietnam activities

= TKI-BBE

= BE-Basic matching

= BE-Basic FES-funding

= Kluyver Centre

= Ecogenomics

= B-Basic

Facts and figures

39

Valorisation output

Size of different funds BE-Basic (M€ per year)

Distribution of BE-Basic funds (%)

BE-BIC and otherSupport office

R&D International projects

R&D total

24

6

94

Flagship 1

Flagship 2

Flagship 3

Flagship 4

Flagship 5

Flagship 6

Flagship 7

Flagship 8

Flagship 9

Flagship 10

Flagship 11

Flagship 12

0 5 10 15 20 25 30Total approved budget per

Flagship (M€)

Nor

th-A

mer

ica

Sout

h-A

mer

ica

Euro

pe

Asi

a

1828

773

150

295

100161186157160

6528210085

106164162123125

4711125

262213 12

210131318

Scientific output

Participants international workshops(2010-2015)

24293

1211594

311010

2119242112

242101

2010/20112012201320142015

Projects for start-ups funded (partly) by BE-Basic

New methods, products, services, protocols and kits developed

Patent filings

Start-ups (i.o.)

Knowledge transfer

Peer reviewed papers

Posters

Presentations total

Contributions to books / book chapters

Non-scientific papers for general public

PhD theses

micro

BE-Basic Foundationaddress Mijnbouwstraat 120

2628 RX DelftThe Netherlands

email info@be-basic.orgweb www.be-basic.orgphone +31 15 2789143