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Industrial Biotechnology – a Key to Bioeconomy

Kirsi-Marja Oksman-CaldenteyVTT Technical Research Centre of Finland

Photo Jouko Lehmuskallio

Polish - Finnish Innovation ForumHelsinki, 8.6.2016

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Biotechnology as an enablerCO2

ENZYME

LIVING CELL

Industrial Biotechnology is a set of practices that usesliving cells or component cells like enzymes, togenerate sustainable industrial products and processes.

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What is possible ?

Ø Biotechnology is suited for large and small scaleØ Production of bioethanol, butanol, lactic acid…Ø Pharmaceutical biotechnology; therapeutic proteins…

Ø Chemical and energy companies turn to biotechnology;DuPont, DSM, BASF, Total, BP

Ø Production levels can be >> 100 g / liter enzymes >> 150g/ liter chemicals, with yields of 30-95%

Ø Production can be improved 10-1000-fold using newtechnologies

Baker’s yeastcell volume ~ 40 µm3

Fermentation tankscan be million liters

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From sugars to fuels and chemicalsEstablished workflow for development of industrial microbes

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Yeasts andbacteria

Filamentousfungi

Algae andcyanobacteria

Plant cells

Biotechnical tools, data analysis, imaging, process development, piloting

Biofuels

Industrial enzymesBioplasticprecursors

Platform andhigh-value chemicals

Therapeuticmolecules

Food and cosmeticingredients

Enzymesin vitro & in vivo

Bionanomaterials

What is our mission?

We develop more efficient and sustainable cell factories, biological molecules andproduction concepts for biotech, pharma, enzyme, energy, chemical and cosmeticindustries

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CASE 1: Novel bacterial strain for ethanol production

Customer: Deinove

Challenge: Unlock the potential of unique bacterial strains forethanol production

Solution: VTT studied the production physiology of Deinococcusbacteria, analysing and characterising the hydrolytic enzymes theyproduced

Key benefits: Set the groundwork for commercialisation of thelignocellulosic ethanol production process 60 fold

improvementin ethanolproduction

“Working with VTT is a great experience, their flexibility andmulti-technology competencies are unmeasurable assets thathelp us speed up our R&D.”

- Jean-Paul Leonetti, VP, R&D Director, Deinove

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CASE 2: Lactic acid - Improved host for bioplasticproduction

Customer: Cargill / NatureWorks LLC

Challenge: To create a new production host for lactic acid productionbetter suited for industrial production conditions

Solution: VTT engineered a yeast strain for more efficient conversion ofplant materials to lactic acid. NatureWorks LLC, subsidiary of Cargill, iscurrently a leading producer of plant based bioplastics.

Key benefit: Contributed to lowering production costs

Bioplastic,world´s leading

Productionlevels up to92 g/l

of lactic acid*

Ref. Ilmén, M. et al. (2013): Microbial Cell Factories 12: 53

”Collaboration with VTT enabled Cargill to explore a larger technologyspace. VTT’s strong expertise in working with non-conventional microbeswas very beneficial in the Cargill project.”

- Pirkko Suominen, AVP, Director Biotechnology Development Center

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CASE 3: Constructing a value chain – Bio-glycolicacid for bio-based polyesters

Ø VTT has developed a value chain from bio-based glycolic acidproduced by fermentation to a value-added polymer

Ø Exploitation potentialØ Barrier material in rigid and flexible packagingØ Total barrier material market > €1billion

914/06/2016 9Decreasing number of candidates

Genetically engineered organisms for theproduction of chemicals

- examples with VTT IPR available for licensing

Xylose

Isoprene

Dicarboxylic acids

Oleochemicals

Xylonic acid

Glycolic acid

Glucose

Pectin

Fatty acids

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VTT Culture Collection http://culturecollection.vtt.fi/

Ø VTT Culture Collection is a fast developing internationally recognisedcollection that provides strains and identification services for the foodand biotechnological sectors worldwide

Ø The total holding of the collection is currently around 6500 microbialstrains and plant cell lines, many are unique strains only available fromVTTCC

> 800 bacterial species from 225 genera> 600 fungal species from 255 generac. 30 plant cell lines from 15 species

Ø Contact:Microbial collections: culture.collection@vtt.fiPlant cell culture collection: plant.collection@vtt.fi

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Enzyme production technology for biorefineriesFrom biomass to sugars

Replace the expensive commercialenzymes with your own production

platform

Ø Trichoderma reesei is a highly efficientprotein production platform (> 100 g/l)

Ø Used in large scale production >100 m3

Ø On-site production concept developmentØ VTT has proprietary production strains

New enzyme discovery andengineering

Optimization of enzyme mixtures fordifferent substrates

Consolidated bioprocessing CBP

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CASE 4: More efficient biomass-to-ethanol production

Customer: Mascoma Corporation

Challenge: Develop a novel production host for more cost-efficient lignocellulosic ethanol production

Solution: VTT engineered a yeast strain capable ofconsolidated bioprocessing (CBP). This significantly reducedthe need for external enzymes, improving the economics ofethanol production.

Key benefit: A more cost-efficient ethanol production method50 %

reduction inthe amountof externalenzymes

”VTT’s deep technical knowledge and great people helped us to deliver onour project. Working with them was truly a great experience!”

- John McBride, Sr. Director R&D, Mascoma Corp.

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Plant cells towards industrial use

Ø We cultivate various plant cell and organ cultures from laboratory to pilot scale(1200 liters) for production of high-value molecules and special proteins

Ø We can also engineer the cells for use as green cell factories in variousindustrial applications (pharma, cosmetics)

Plant

Callus Cell suspension

Cells in bioreactor

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Scaling up the cloudberry cells to bioreactors anddownstream processing

300 L steel tankbioreactor

2 – 50 L disposablewave bioreactor

Filter press Lyophilisation

Raw materialfor the finalproduct

Nohynek, L., Bailey, M., Tähtiharju, J., Seppänen-Laakso, T., Rischer, H., Oksman-Caldentey, K.-M. & Puupponen-Pimiä, R. (2014): Cloudberry (Rubus chamaemorus) cellculture with bioactive substances: establishment and mass propagation for industrial use.Eng Life Sci 14: 667-675.

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CASE 5: Sustainable cosmetics from cloudberry cellsCustomer: Lumene

Challenge: Enable the sustainable production of bioactivecompounds from a natural source

Solution: The VTT Plant biotechnology team established asingle cell culture from a rare cloudberry plant enabling thesustainable production of bioactive compounds in controlledbioreactor environment

Key benefits: Leveraging the bioactivities of cloudberry stemcell culture enabled the creation of a new line of cosmetics new line

of cosmeticsfrom

cloudberrystem cells

“Achieving international growth in the cosmetics industry requires notonly solid innovative product development, but also good, world classpartners. VTT is a natural and preferred partner for us, because theyhave innovative expertise and dedication to succeed in the utilization ofArctic berries with the latest technology.”

- Tiina Isohanni, VP Innovations and Development, Lumene

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Parallel small scale bioreactor systems(0.2 L – 1.0 L) for strain evaluation

Cost efficient screening in bioreactor environment

Up- and downstream process optimization (2 L – 20 L)

Possibilities for transcriptome and flux balance analysisOn-line analytics for efficient process control

Process scale-up (300 L, 1200 L)

Confirmation of process performance in fully equippedbiomanufacturing pilot plant

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Tailored living cell factories –the whole value chain for the benefit of your company

Gene/enzyme discovery and engineering ® metabolic/protein design ® hoststrain development ® validation ® process concepts from lab to pilot ® up-scaling and down stream processing ® final product

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Wihuri International Awardto Merja Penttilä

2012

Industrial BiotechnologyOVER 30 YEARS FROM SCIENCE TO INNOVATIONS

1985 1990 1995 2000 2005 2010 2015 2020

19804 p-years

199015 p-years

200045 p-years

201385 p-years

2016>90 p-years

VTT Industrial BiotechnologyCentre of Excellence

2000-2005

1980 gene technology1985 metabolic engineering1985 plant biotechnology

Trichodermatechnology

transfer to Alko

Trichodermatransformation

&cellulases

White biotechnology –Green Chemistry Research

Centre of Excellence2008-2013

NatureWorksLactic acid

Xylitol yeastPentose yeast

SolucelSpin-off

Holistic approachto cell factories

Genencor

MascomaLignocellulosic

ethanol

New products:energy carriers,

bio-monomers, high-value and platform

chemicals

LumeneBerry cell technology

Development ofGM barley

for PBL & Alko

Glykos/NovartisTherapeutic

proteins

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Why partner with VTT Industrial Biotechnology?

Ø One-stop-shop: Strain and process development, bioinformatics, enzymology,analytics, techno-economics, patent surveys – all in one project team

Ø More than 100 people with broad expertise background in industrial biotechnology

Ø Excellent research infrastructure from lab to pilot scale

Ø Over 30 years of experience working with the industry

VTT delivers results!

Technology for business