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Industrial Biotechnology – a Key to Bioeconomy
Kirsi-Marja Oksman-Caldentey
VTT Technical Research Centre of Finland
Photo Jouko Lehmuskallio
Polish - Finnish Innovation Forum
Helsinki, 8.6.2016
214/06/2016 2
Biotechnology as an enablerCO2
ENZYME
LIVING CELL
Industrial Biotechnology is a set of practices that uses
living cells or component cells like enzymes, to
generate sustainable industrial products and processes.
314/06/2016 3
What is possible ?
Ø Biotechnology is suited for large and small scale
Ø 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 >> 150
g/ liter chemicals, with yields of 30-95%
Ø Production can be improved 10-1000-fold using new
technologies
Baker’s yeast
cell volume ~ 40 µm3
Fermentation tanks
can be million liters
414/06/2016 4
From sugars to fuels and chemicals
Established workflow for development of industrial microbes
5
Yeasts and
bacteria
Filamentous
fungi
Algae and
cyanobacteria
Plant cells
Biotechnical tools, data analysis, imaging, process development, piloting
Biofuels
Industrial enzymes
Bioplastic
precursors
Platform and
high-value chemicals
Therapeutic
molecules
Food and cosmetic
ingredients
Enzymes
in vitro & in vivo
Bionanomaterials
What is our mission?
We develop more efficient and sustainable cell factories, biological molecules and
production concepts for biotech, pharma, enzyme, energy, chemical and cosmetic
industries
6614/06/2016
CASE 1: Novel bacterial strain for ethanol production
Customer: Deinove
Challenge: Unlock the potential of unique bacterial strains for
ethanol production
Solution: VTT studied the production physiology of Deinococcus
bacteria, analysing and characterising the hydrolytic enzymes they
produced
Key benefits: Set the groundwork for commercialisation of the
lignocellulosic ethanol production process
60 fold
improvement
in ethanol
production
“Working with VTT is a great experience, their flexibility and
multi-technology competencies are unmeasurable assets that
help us speed up our R&D.”
- Jean-Paul Leonetti, VP, R&D Director, Deinove
7714/06/2016
CASE 2: Lactic acid - Improved host for bioplastic
production
Customer: Cargill / NatureWorks LLC
Challenge: To create a new production host for lactic acid production
better suited for industrial production conditions
Solution: VTT engineered a yeast strain for more efficient conversion of
plant materials to lactic acid. NatureWorks LLC, subsidiary of Cargill, is
currently a leading producer of plant based bioplastics.
Key benefit: Contributed to lowering production costs
Bioplastic,
world´s leading
Production
levels up to
92 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 technology
space. VTT’s strong expertise in working with non-conventional microbes
was very beneficial in the Cargill project.”
- Pirkko Suominen, AVP, Director Biotechnology Development Center
8
CASE 3: Constructing a value chain – Bio-glycolic
acid for bio-based polyesters
Ø VTT has developed a value chain from bio-based glycolic acid
produced by fermentation to a value-added polymer
Ø Exploitation potential
Ø Barrier material in rigid and flexible packaging
Ø Total barrier material market > €1billion
914/06/2016 9
Decreasing number of candidates
Genetically engineered organisms for the
production of chemicals
- examples with VTT IPR available for licensing
Xylose
Isoprene
Dicarboxylic acids
Oleochemicals
Xylonic acid
Glycolic acid
Glucose
Pectin
Fatty acids
1014/06/2016 10
VTT Culture Collection http://culturecollection.vtt.fi/
Ø VTT Culture Collection is a fast developing internationally recognised
collection that provides strains and identification services for the food
and biotechnological sectors worldwide
Ø The total holding of the collection is currently around 6500 microbial
strains and plant cell lines, many are unique strains only available from
VTTCC
> 800 bacterial species from 225 genera
> 600 fungal species from 255 genera
c. 30 plant cell lines from 15 species
Ø Contact:
Microbial collections: culture.collection@vtt.fi
Plant cell culture collection: plant.collection@vtt.fi
1114/06/2016 11
Enzyme production technology for biorefineries
From biomass to sugars
Replace the expensive commercial
enzymes with your own production
platform
Ø Trichoderma reesei is a highly efficient
protein production platform (> 100 g/l)
Ø Used in large scale production >100 m3
Ø On-site production concept development
Ø VTT has proprietary production strains
New enzyme discovery and
engineering
Optimization of enzyme mixtures for
different substrates
Consolidated bioprocessing CBP
121214/06/2016
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 of
consolidated bioprocessing (CBP). This significantly reduced
the need for external enzymes, improving the economics of
ethanol production.
Key benefit: A more cost-efficient ethanol production method
50 %
reduction in
the amount
of external
enzymes
”VTT’s deep technical knowledge and great people helped us to deliver on
our project. Working with them was truly a great experience!”
- John McBride, Sr. Director R&D, Mascoma Corp.
13
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 various
industrial applications (pharma, cosmetics)
Plant
Callus Cell suspension
Cells in bioreactor
14
Scaling up the cloudberry cells to bioreactors and
downstream processing
300 L steel tank
bioreactor
2 – 50 L disposable
wave bioreactor
Filter press Lyophilisation
Raw material
for the final
product
Nohynek, L., Bailey, M., Tähtiharju, J., Seppänen-Laakso, T., Rischer, H., Oksman-
Caldentey, K.-M. & Puupponen-Pimiä, R. (2014): Cloudberry (Rubus chamaemorus) cell
culture with bioactive substances: establishment and mass propagation for industrial use.
Eng Life Sci 14: 667-675.
151514/06/2016
CASE 5: Sustainable cosmetics from cloudberry cells
Customer: Lumene
Challenge: Enable the sustainable production of bioactive
compounds from a natural source
Solution: The VTT Plant biotechnology team established a
single cell culture from a rare cloudberry plant enabling the
sustainable production of bioactive compounds in controlled
bioreactor environment
Key benefits: Leveraging the bioactivities of cloudberry stem
cell culture enabled the creation of a new line of cosmetics
new line
of cosmetics
from
cloudberry
stem cells
“Achieving international growth in the cosmetics industry requires not
only solid innovative product development, but also good, world class
partners. VTT is a natural and preferred partner for us, because they
have innovative expertise and dedication to succeed in the utilization of
Arctic berries with the latest technology.”
- Tiina Isohanni, VP Innovations and Development, Lumene
1614/06/2016 16
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 analysis
On-line analytics for efficient process control
Process scale-up (300 L, 1200 L)
Confirmation of process performance in fully equipped
biomanufacturing pilot plant
17
Tailored living cell factories –
the whole value chain for the benefit of your company
Gene/enzyme discovery and engineering ® metabolic/protein design ® host
strain development ® validation ® process concepts from lab to pilot ® up-
scaling and down stream processing ® final product
1814/06/2016 18
Wihuri International Award
to Merja Penttilä
2012
Industrial Biotechnology
OVER 30 YEARS FROM SCIENCE TO INNOVATIONS
1985 1990 1995 2000 2005 2010 2015 2020
1980
4 p-years
1990
15 p-years
2000
45 p-years
2013
85 p-years
2016
>90 p-years
VTT Industrial Biotechnology
Centre of Excellence
2000-2005
1980 gene technology
1985 metabolic engineering
1985 plant biotechnology
Trichoderma
technology
transfer to Alko
Trichoderma
transformation
&
cellulases
White biotechnology –
Green Chemistry Research
Centre of Excellence
2008-2013
NatureWorks
Lactic acid
Xylitol yeast
Pentose yeast
Solucel
Spin-off
Holistic approach
to cell factories
Genencor
Mascoma
Lignocellulosic
ethanol
New products:
energy carriers,
bio-monomers, high-
value and platform
chemicals
Lumene
Berry cell technology
Development of
GM barley
for PBL & Alko
Glykos/Novartis
Therapeutic
proteins
14/06/2016 19
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

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Industrial Biotechnology-a Key to Bioeconomy

  • 1. 11 Industrial Biotechnology – a Key to Bioeconomy Kirsi-Marja Oksman-Caldentey VTT Technical Research Centre of Finland Photo Jouko Lehmuskallio Polish - Finnish Innovation Forum Helsinki, 8.6.2016
  • 2. 214/06/2016 2 Biotechnology as an enablerCO2 ENZYME LIVING CELL Industrial Biotechnology is a set of practices that uses living cells or component cells like enzymes, to generate sustainable industrial products and processes.
  • 3. 314/06/2016 3 What is possible ? Ø Biotechnology is suited for large and small scale Ø 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 >> 150 g/ liter chemicals, with yields of 30-95% Ø Production can be improved 10-1000-fold using new technologies Baker’s yeast cell volume ~ 40 µm3 Fermentation tanks can be million liters
  • 4. 414/06/2016 4 From sugars to fuels and chemicals Established workflow for development of industrial microbes
  • 5. 5 Yeasts and bacteria Filamentous fungi Algae and cyanobacteria Plant cells Biotechnical tools, data analysis, imaging, process development, piloting Biofuels Industrial enzymes Bioplastic precursors Platform and high-value chemicals Therapeutic molecules Food and cosmetic ingredients Enzymes in vitro & in vivo Bionanomaterials What is our mission? We develop more efficient and sustainable cell factories, biological molecules and production concepts for biotech, pharma, enzyme, energy, chemical and cosmetic industries
  • 6. 6614/06/2016 CASE 1: Novel bacterial strain for ethanol production Customer: Deinove Challenge: Unlock the potential of unique bacterial strains for ethanol production Solution: VTT studied the production physiology of Deinococcus bacteria, analysing and characterising the hydrolytic enzymes they produced Key benefits: Set the groundwork for commercialisation of the lignocellulosic ethanol production process 60 fold improvement in ethanol production “Working with VTT is a great experience, their flexibility and multi-technology competencies are unmeasurable assets that help us speed up our R&D.” - Jean-Paul Leonetti, VP, R&D Director, Deinove
  • 7. 7714/06/2016 CASE 2: Lactic acid - Improved host for bioplastic production Customer: Cargill / NatureWorks LLC Challenge: To create a new production host for lactic acid production better suited for industrial production conditions Solution: VTT engineered a yeast strain for more efficient conversion of plant materials to lactic acid. NatureWorks LLC, subsidiary of Cargill, is currently a leading producer of plant based bioplastics. Key benefit: Contributed to lowering production costs Bioplastic, world´s leading Production levels up to 92 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 technology space. VTT’s strong expertise in working with non-conventional microbes was very beneficial in the Cargill project.” - Pirkko Suominen, AVP, Director Biotechnology Development Center
  • 8. 8 CASE 3: Constructing a value chain – Bio-glycolic acid for bio-based polyesters Ø VTT has developed a value chain from bio-based glycolic acid produced by fermentation to a value-added polymer Ø Exploitation potential Ø Barrier material in rigid and flexible packaging Ø Total barrier material market > €1billion
  • 9. 914/06/2016 9 Decreasing number of candidates Genetically engineered organisms for the production of chemicals - examples with VTT IPR available for licensing Xylose Isoprene Dicarboxylic acids Oleochemicals Xylonic acid Glycolic acid Glucose Pectin Fatty acids
  • 10. 1014/06/2016 10 VTT Culture Collection http://culturecollection.vtt.fi/ Ø VTT Culture Collection is a fast developing internationally recognised collection that provides strains and identification services for the food and biotechnological sectors worldwide Ø The total holding of the collection is currently around 6500 microbial strains and plant cell lines, many are unique strains only available from VTTCC > 800 bacterial species from 225 genera > 600 fungal species from 255 genera c. 30 plant cell lines from 15 species Ø Contact: Microbial collections: culture.collection@vtt.fi Plant cell culture collection: plant.collection@vtt.fi
  • 11. 1114/06/2016 11 Enzyme production technology for biorefineries From biomass to sugars Replace the expensive commercial enzymes with your own production platform Ø Trichoderma reesei is a highly efficient protein production platform (> 100 g/l) Ø Used in large scale production >100 m3 Ø On-site production concept development Ø VTT has proprietary production strains New enzyme discovery and engineering Optimization of enzyme mixtures for different substrates Consolidated bioprocessing CBP
  • 12. 121214/06/2016 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 of consolidated bioprocessing (CBP). This significantly reduced the need for external enzymes, improving the economics of ethanol production. Key benefit: A more cost-efficient ethanol production method 50 % reduction in the amount of external enzymes ”VTT’s deep technical knowledge and great people helped us to deliver on our project. Working with them was truly a great experience!” - John McBride, Sr. Director R&D, Mascoma Corp.
  • 13. 13 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 various industrial applications (pharma, cosmetics) Plant Callus Cell suspension Cells in bioreactor
  • 14. 14 Scaling up the cloudberry cells to bioreactors and downstream processing 300 L steel tank bioreactor 2 – 50 L disposable wave bioreactor Filter press Lyophilisation Raw material for the final product Nohynek, L., Bailey, M., Tähtiharju, J., Seppänen-Laakso, T., Rischer, H., Oksman- Caldentey, K.-M. & Puupponen-Pimiä, R. (2014): Cloudberry (Rubus chamaemorus) cell culture with bioactive substances: establishment and mass propagation for industrial use. Eng Life Sci 14: 667-675.
  • 15. 151514/06/2016 CASE 5: Sustainable cosmetics from cloudberry cells Customer: Lumene Challenge: Enable the sustainable production of bioactive compounds from a natural source Solution: The VTT Plant biotechnology team established a single cell culture from a rare cloudberry plant enabling the sustainable production of bioactive compounds in controlled bioreactor environment Key benefits: Leveraging the bioactivities of cloudberry stem cell culture enabled the creation of a new line of cosmetics new line of cosmetics from cloudberry stem cells “Achieving international growth in the cosmetics industry requires not only solid innovative product development, but also good, world class partners. VTT is a natural and preferred partner for us, because they have innovative expertise and dedication to succeed in the utilization of Arctic berries with the latest technology.” - Tiina Isohanni, VP Innovations and Development, Lumene
  • 16. 1614/06/2016 16 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 analysis On-line analytics for efficient process control Process scale-up (300 L, 1200 L) Confirmation of process performance in fully equipped biomanufacturing pilot plant
  • 17. 17 Tailored living cell factories – the whole value chain for the benefit of your company Gene/enzyme discovery and engineering ® metabolic/protein design ® host strain development ® validation ® process concepts from lab to pilot ® up- scaling and down stream processing ® final product
  • 18. 1814/06/2016 18 Wihuri International Award to Merja Penttilä 2012 Industrial Biotechnology OVER 30 YEARS FROM SCIENCE TO INNOVATIONS 1985 1990 1995 2000 2005 2010 2015 2020 1980 4 p-years 1990 15 p-years 2000 45 p-years 2013 85 p-years 2016 >90 p-years VTT Industrial Biotechnology Centre of Excellence 2000-2005 1980 gene technology 1985 metabolic engineering 1985 plant biotechnology Trichoderma technology transfer to Alko Trichoderma transformation & cellulases White biotechnology – Green Chemistry Research Centre of Excellence 2008-2013 NatureWorks Lactic acid Xylitol yeast Pentose yeast Solucel Spin-off Holistic approach to cell factories Genencor Mascoma Lignocellulosic ethanol New products: energy carriers, bio-monomers, high- value and platform chemicals Lumene Berry cell technology Development of GM barley for PBL & Alko Glykos/Novartis Therapeutic proteins
  • 19. 14/06/2016 19 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!