EMPOWERING BIOINNOVATION BIO PACIFIC RIM SAN DIEGO 12/9/2014 NAGIB M. WARD, EXECUTIVE VP
DEINOVE CREATES RENEWABLE, RESPONSIBLE AND COST-EFFECTIVE INDUSTRIAL PROCESSES OUR AMBITION Replace petroleum-based molecules with bio-based alternatives OUR CORE ACTIVITY Develop innovative bioproduction processes leveraging the exceptional properties of Deinococcus bacteria OUR COMPETITIVE ADVANTAGES A unique proprietary bacterium, source of key industrial benefits Powerful tool in metabolic engineering Cost-effective and Consolidated BioProcess
DEINOVE 6,000+ STRAINS LIBRARY All strains were collected according to international laws (CBD) Strategically, diverse environments were targeted with: unusual selection by irradiation unusual carbon sources An original and new source of biocatalysts!
Number of publications Number of patents THE GENUS DEINOCOCCUS A NEW SCIENTIFIC AND TECHNOLOGICAL OPPORTUNITY NUMBER OF PUBLICATIONS PER GENUS NUMBER OF PATENTS PER GENUS 300 000 8 000 200 000 100 000 4 000 Escherichia coli Saccharomyces cerevisiae Deinococcus Deinococcus geothermalis Microorganisms 0 Microorganisms The largest collection of Deinos in the World (6,000 strains) A proprietary technological platform (16 patents) with validation through a POC (DEINOL) The World s pioneer in Deino-based technologies and their IP.
DEINOVE COMMERCIAL STRUCTURE Deinol Deinochem Deinobiotics Green Solvents Biofuels Renewable Chemicals Intermediates Deinomediation ThanaPlast TM
Advanced BioFuels Nutraceuticals Green Fuel Equivalents Cellulosic Ethanol Green Chemicals IsoPropanol PDO C5 Sugars C6 Sugars C5 Sugars C6 Sugars C5 Sugars C6 Sugars Glyceraldehyde-3P Glycerol Deinol Deinochem Cellulosic Biomass Starch Cellulosic Biomass Starch Saponification CoP 1 st Gen Fiber 1 st Gen Fiber Biodiesel CoP MSW Organics
A TECHNOLOGY PROVIDER VIA LICENSING SOLUTIONS & BUSINESS MODEL 3: Feedstock Our strains are industrial fermentation hosts, with CBP capability. Dedicated fermentations with feedstock flexibility, and process cost benefits. 4: Process Benefits High-value Carotenoids available from the bacterial mass, extractable postfermentation. 1: Main Product 2: Co-Product
DEINOVE FERMENTATION LIBRARY PIGMENTED BACTERIA BIODIVERSITY: PINK, ORANGE AND RED Deinococcus ficus Deinococcus aquaticus Deinococcus daejeonensis Deinococcus marmoris Roseomonas aestuarii Rhodococcus corynebacterioides Kocuria turfanensis
DEINOVE FERMENTATION LIBRARY A WIDE VARIETY OF PIGMENTS
Mx6-1B DRH-46 M1-3H M13-1A MC3-9A MI3-1C M11-12B MI3_10 M23r-2A M23-2E M31-8F MC5-11A MX4-5F M31-10 M36-8C M19-GG M26-8A MC3-4A MC3-4B MC3-5G MC2-2C MC2-2A M36-70 MX6-1C MX6-10 MX6-1E DRH-37 DRH-38 DSM11300 BIOMASS HYDROLYZING ENZYMES 50 STRAINS SEQUENCED, PHYLOGENY OF GLYCOSYL HYDROLASES GH13 GH13 GH13 GH13 GH13 GH13 GH13 GH13 GH77 GH13 GH15 GH13 GH13 GH42 GH36 GH1 GH101 GH8 GH52 GH5 GH3 GH78 GH3 5.4.99.15 3.2.1.141 2.4.1.18 2.4.1.25 (33) 2.4.1.4 3.2.1.68 3.2.1.20 3.2.1.1 2.4.1.25 3.2.1.20(28) 3.2.1.3 3.2.1.1 3.2.1.41(6 3.2.1.23 8) 3.2.1.22 3.2.1.21 23.2.1.97 3.2.1.8 3.2.1.37 3.2.1.4 3.2.1.215 3.2.1.40 2) 3.2.1 Maltooligosyl trehalose synthase Maltooligosyl trehalose trealohydrase 1,4 aglucan branching enzyme 4 aglucanotransferase; amiloa 1,6 glucosidase Amylosucrase Isoamylase aglucosidase aamylase 4agluconotransferase aglucosidase; atrehalase Glucoamylase aamylase Pellulanase; Isoamylase bgalactosidase agalactosidase bglucosidase GlycopeptideaN acetylgalactosaminidase Endo 1,4 bxylanase Xylane 1,4 bxylosidase Cellulase bglucosidase; bnacetylhexosaminidase alrhamnosidase XYLANOLYTIC CELLULOLYTIC AMYLOLYTIC
DEINOVE SOLUTIONS THREE SYNERGISTIC COMPONENTS First: A one-a-kind bacterial library, primary focused on the Deinococcus Genus. Strains and their Properties Second: Deinove s unique and proprietary genetics and metabolism technology to biotransform its organisms into fermentation hosts. Third: A fermentation process savoir-faire for rare fermentation hosts. Process Integration & Partner Transfer Molecular Biology and Metabolomics
DEINOL
Temperature
Molecular Biology/Metabolomics
DEINOVE BIOTRANSFORMATION Specific molecular biology tools are necessary to biotransform Deinove s strains. Exact location for genetic insertions, or deletions (KO). Numerous genetic cassettes can be expressed at high levels.
new sites in progress Available sites 0 Multi-targetting cassette Specific targetting cassette 315 45 270 Deinococcus main chromosome 90 225 135 180
Unique Inherent Properties
THE DEINOCOCCUS ADVANTAGES INHIBITORS RESISTANCE Metabolization test (MTT) IC 50 (g/l) 5-HMF Furfural Aldehyde from lignin E. coli 26 3 5 S. cerevisiae 18 1 3 D. radiodurans R1 93 20 14 D. geothermalis MX6-1E 104 42 12 D. geothermalis MX6-1C 112 46 9 D. geothermalis MC2-2A 112 3 13 D. geothermalis M36-7D 97 30 ND
%total HEMICELLULOLYSIS ASSIMILATION OF A MIX OF BIRCH XYLAN 120 100 Kinetic of xylose assimilation on birch wood xylan 80% of the total xylan is consumed after 48h of cultivation. 80 60 C5 Xylo-oligomers are preferably absorbed, and metabolized intracellularly with beta-xylosidase. 40 20 0 0 20 40 60 80 100 120 140 160 180
g/l g/l g/l g/l CELLULOLYSIS ASSIMILATION OF CELLO-OLIGOMERS 16 14 12 10 8 6 4 2 0 0 10 GLUCOSE 20 30 40 50 60 70 80 16 14 12 10 8 6 4 2 0 0 10 CELLOTRIOSE 20 30 40 50 60 70 80 Time(h) Time(h) 16 14 12 10 8 6 4 2 0 0 10 CELLOBIOSE 20 30 40 50 60 70 80 16 14 12 10 8 6 4 2 0 0 10 CELLOTETRAOSE 20 30 40 50 60 70 80 Time(h) Time(h)
All Sugars without diauxie. Really.
Total sugar consumed g Xylose consumed g Glucose consumed g EtOH produced g 6000 5000 4000 3000 2000 1000 0 600 500 400 300 200 100 0 6000 5000 4000 3000 2000 1000 0 1 2 1 2 0 24 48 72 96 120 144 168 192 Time h 0 24 48 72 96 120 144 168 192 Time h 0 24 48 72 96 120 144 168 192 Time h 350 300 250 200 150 100 50 0 propagation 0 24 48 72 96 120 144 168 192 Time h Green line are kinetic predicted values production Red points and lines are measured values Glucose and xylose are co-assimilated without diauxie.
G/L SUGAR ASSIMILATION 20 18 16 14 12 10 8 6 4 2 0 XYLAN CELLOBIOSE 0 10 20 30 40 50 60 HOURS
Consolidated BioProcess
DEINOL BENEFITS PROCESS CONSIDERATIONS SSF Hydrolysis Biomass Pretreatment Amorphous Cellulose and Hemicellulose Enzymes Cello-Oligomers Xylo-Oligomers Deinos Enzymes 1. Main Product 2. Coproduct 3. Energy Downstream Processing Fermentation GM-Non CBP Yeast C6/C5 Monosaccharides
DEINOVE ENZYME PORTFOLIO SIMPLIFICATION Cellulose Polymeric: Exoglucanases (CBHs) Endoglucanases (Egs) Glucosyl Hydrolases (GH61) Saccharification: Beta-Glucosidase (BG) Hemicellulose Polymeric: Exoxylanases Endoxylanases Saccharification: Beta-Xylosidase (Bx) Acetyl Xylan Esterase (AXE) Ferulic Acid Esterase (FAE)
PROCESS PROFITABILITY ADVANTAGES CBP ENZYME BOOSTING The fermentation temperature preference range of 45-50C is ideal: it leverages the biochemical temperature optimum for both intracellular and exogenous cellulosic enzymes. Cost benefit: ~30%. Xylo-oligomers (known endoglucanase inhibitors), and cello-oligomers are preferably absorbed and metabolized, thereby minimizing both the exogenous cellulase and hemicellulase doses required. Cost Benefit: ~30% AXE, Bg and Bx are no longer required in solution, which significantly simplify the exogenous enzyme product composition necessary and improves the producer s production rates. Cost Benefit: ~20% An estimated combined 50 to 65% enzyme cost reduction, while providing additional variable and capex cost reduction opportunities.
Biomass Pretreatment Amorphous Cellulose and Hemicellulose Enzymes Cello-Oligomers Xylo-Oligomers Ethanol Downstream Processing Deinos Terpenoids Advanced Biofuel Downstream Processing Fermentation Chemical Intermediate
CO-DEVELOPMENT BUSINESS MODEL TECHNOLOGY PROVIDER VIA LICENSING Confidentiality NDA Material transfer MTA Preliminary tests or POC Collaborative agreement *Non exclusive or Limited Exclusivity JDA* Scale-up and Integration INDUSTRIALIZATION COMMERCIALIZATION
CONTACTS Nagib M. WARD Vice President, Business Development nagib.ward@deinove.com CapTrust Tower, 4208 Six Forks Road, Suite 1000 Raleigh, NC 27609 USA (O): +1 919-645-5784 (M): +1 609-772-2223 (F): +1 919-825-4801 Jean-Paul LEONETTI VP R&D Tel : + 33 1 42 03 27 37 jean-paul.leonetti@deinove.com Emmanuel PETIOT CEO Tel : + 33 1 42 03 27 37 emmanuel.petiot@deinove.com
Background Slides
DEINOVE INVESTMENT HIGHLIGHTS DISRUPTIVE TECHNOLOGY Access to biomass carbon, substituting for enzymes+yeasts Lower production costs and performance improvement thanks to thermophilic properties Consolidated bioprocess (CBP) and metabolic activities allowing higher productivity and yield VERSATILE TECHNOLOGY Wide range of Deinococcus strains offering endless applications Core biological process may address any kind of input substrate and output compound RELEVANT PARTNERS LONG-TERM MARKET POTENTIAL Abengoa, a leading bioethanol producer Suez Environnement to develop waste-to-ethanol Urge to reduce dependence on expensive fossil energies Lignocellulosic feedstock avoid competition for agricultural land while population is growing fast Political support for developing biofuels and biochemicals Opportunity to substantially reduce carbon footprint
DEINOCOCCUS BACTERIA: A NOVEL HOST POTENTIAL TO EXCEED KNOWN LIMITATIONS OF KNOWN STRAINS DEINOCOCCUS PROPERTIES INDUSTRIAL ADVANTAGES EXCEPTIONAL ROBUSTNESS Furfural/HMF/aldehydes, organic acids GREAT ABILITY TO DEGRADE BIOMASS Intrinsic cellulolytic and hemicellulolytic activities RELIABLE BACTERIAL ACTIVITY ALONG THE PROCESS REDUCED PRODUCTION COSTS (ENZYMES, CAPEX) AWARDED MICROBE OF THE YEAR 2014 (DEMO. SCALE) BY BIOFUELS DIGEST FERMENTATION Simultaneous assimilation of C6&C5 sugars HIGH GENOMIC STABILITY Robust genetic modifications THERMOPHILIC Operating range 45 C to 50 C NATURAL EXPRESSION OF CAROTENOIDS NON-PATHOGENIC For humans, animals and environment HIGH-YIELD PROCESS STABLE AND REPRODUCIBLE PRODUCTION SYSTEMS REDUCED ENERGY CONSUMPTION RARE OR TECHNOLOGICALLY INACCESSIBLE PRODUCTS EASY TO UPGRADE EXISTING FACILITIES