PRESENTATION BIO WORLD CONGRESS 19 JUNE 2013

Transcription

1 PRESENTATION BIO WORLD CONGRESS 19 JUNE 2013

2 COMPANY SNAPSHOT DEINOVE, THE DEINOCOCCUS WAY DEINOVE is a cleantech company that designs, develops and markets a new generation of industrial processes based on ancestral bacteria with untapped potential: the deinococci. These new systems open the way for manufacturing of rare or technologically inaccessible products: Second-generation biofuels (DEINOL) High added value biochemicals (DEINOCHEM) New antibiotics (DEINOBIOTICS) Enzymes for the remediation of plastics (THANAPLAST ) DEINOVE intends to provide new industrial technology standards that enable it to meet the great world challenges: preparing for the post-oil era, switching to green chemistry, fighting against antibiotic resistance and improving the life cycle of plastics. 2

3 DEINOVE KEY DATES Proprietary technological platform Initiation of DEINOBIOTICS program Initiation of DEINOCHEM program DEINOBIOTICS SAS Deinobank (25 strains) Deinobank (6 000 strains) Initiation of DEINOL program DEINOL EC2 (POC) THANAPLAST program DEINOL EC Inception, co-founders Pr Miroslav RADMAN Dr Philippe POULETTY OSEO funding ( 6 m) OSEO funding ( 1.57 m) Emmanuel PETIOT CEO Jacques BITON CEO IPO ( 12 m) Partnership with TEREOS Partnership with VTT OSEO funding ( 1.4 m) OSEO funding ( 1.15 m) Market Cap: 53 m Staff

4 DEINOVE STRATEGY FROM 2012 to 2013 Second generation biofuels Other programs Plastic remediation Antibiotics = DEINOL and beyond THANAPLAST DEINOBIOTICS Plastics Fine chemicals Sweeteners Fragrances Flavors Detergents DEINOCHEM 4

5 DEINOCOCCUS: A NEW CHASSIS FOR FERMENTATION 5

6 DEINOCOCCUS: A NEW CHASSIS FOR 2 nd GENERATION BIOFUELS A UNIQUE SUM OF BENEFITS D. geothermalis S. cerevisiae E. coli C. glutamicum C5 and C6 assimilation C5 and C6 co-assimilation amylolytic cellulolytic Cello (bi-tri-tetra) ose Xylanolytic Resistance to dessication Resistance to xenobiotics Rs glucose Biomass in a mineral media Kla Thermophile Genetic engineering tools 6

7 Number of publications Number of patents THE GENUS DEINOCOCCUS A NEW SCIENTIFIC AND TECHNOLOGICAL WAY NUMBER OF PUBLICATIONS PER GENUS NUMBER OF PATENTS PER GENUS Escherichia coli Saccharomyces cerevisiae Deinococcus Deinococcus geothermalis Microorganisms 0 Microorganisms The largest collection of Deinococci in the world (6 000 strains) A proprietary technological platform (16 patents) with validation through a POC (DEINOL) The world pioneer of deinotechnologies 7

8 OUR PROPRIETARY TECHNOLOGICAL PLATFORM THE BEST OF GENETIC ENGINEERING AND BACTERIOLOGY COLLECTION ARCHIVING SELECTION OPTIMISATION SCALE UP Samples collection in extreme environment Library of cryopreserved strains Physiology High-throughput growth assays Metabolomic Functional tests Gene expression measurement Microbiology Genomic PCR Sequencing & Annotation Genetic & Metabolic Engineering Gene insertion Gene deletion Sequence optimization Fermentation engineering Optimization of culture & fermentation processes Industrial engineering Lab Assays Test & Integration DEINOBANK (6 000 clones) S1 S2 S3 LEAD DEINOCOCCUS POC CANDIDAT LAB PILOT INDUSTRIAL PILOT PRODUCTION 8

9 THE DEINOVE TEAM A YOUNG INNOVATIVE COMPANY 9

10 A BACTERIAL LIBRARY OF STRAINS MANY UNSUAL STRAINS DUE TO SELECTION BY IRRADIATION Diverse environments have been targeted unusual selection by irradiation unusual carbon sources Deinococcus library (1800 strains) AND diverse library of original strains (4200) All strains collected according to the international laws (CBD) Source of original new biocatalysts 10

11 THE DEINOCOCCUS ADVANTAGES INHIBITORS RESISTANCE Metabolization test (MTT) IC 50 (g/l) 5-HMF Furfural Aldehyde from lignin E. coli S. cerevisiae D. radiodurans R D. geothermalis MX6-1E D. geothermalis MX6-1C D. geothermalis MC2-2A D. geothermalis M36-7D ND 11

12 THE DEINOCOCCUS ADVANTAGES A GENETIC MOSAIC WITH UNUSUAL ROBUSTNESS Among the most resistant genomes of the world : Most of the publications on D. radiodurans (900) focused on radiation resistance Multiple copies of the genome (Battista J, Annu. Rev. Microbiol. 1997) Specific DNA repair machinery (Repar J, DNA Repair 2010) ETOH OPERON BIOMASS OPERON Recombinant D. geothermalis 2 Recombinant D. geothermalis after serial dilution 3 Wild type D. geothermalis, 2 synthetic operons have been inserted and the bacteria have been diluted without selection pressure. Both are stable after 100 generations even without selection pressure 12

13 OD 570 NM (% CONTROL) THE DEINOCOCCUS ADVANTAGES NOW ACCESSIBLE TO DIRECTED EVOLUTION E. coli S. Cerevisiae D. radiodurans R1 Recombinant D. geothermalis Evolved recombinant D. geothermalis %% ETHANOL ÉTHANOL 13

14 DEINOVE S PLATFORM 14

15 PLATFORM STRAIN CONSTRUCTION PLATFORM A large strain library 6000 strains, 1800 Deinococcus Omic databases genomic, transcriptomic, proteomic Library of standard elements: promotors RBS A robotic platform Generated strains Rémi Bernard Molecular Biology Manager Robotic cloning Dedicated software S S S S Deinococcus allows fast strain construction 15

16 PLATFORM FERMENTATION METABOLISM ANALYTIC Fermentation & Metabolism Analytical Platform Multi parallel fermentors: 1L, 2 L, 5L, 20L. Enzymology, fluxomic, metabolomic MicrOTOF-QII Bruker (intra and extracellular metabolites) Gaz chromatorgraphy variant Dionex ICS (sugars, acids) 16

17 DEINOCOCCUS ENZYMES, BIOMASS DEGRADATION, SUGAR ASSIMILATION 17

18 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 GH (33) (28) (68) ) 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 18

19 BIOMASS HYDROLYZING ENZYMES DEINOCOCCUS ENDOCELLULASE QC SH Cellulase SpF CBM 1728 CBM 1728 Cellulase superfamily Closest relative: Clostridium cellulolyticum GH5 b1,4-glucanase Clostridium cellulolyticum (65% identity) Deinococcus Control (no bacteria) AZO-CMC (cellulose 0.5%) 19

20 BIOMASS HYDROLYZING ENZYMES GH10 XYLANASE QC SH SpF Glyco hydro 10 Glyco hydro 10 superfamily Glycoside hydrolase family 10, closest relative Clostridium thermocellum (45% identity) AZO-Xylan (0.5%) 20

21 Free sugars (g/l) DEINOCCOCUS IN THE REAL LIFE ASSIMILATION OF MIXES OF GLUCOSE & XYLOSE % OF RS MAX WHEAT BRAN, shake flask Ratio glucose/xylose g/g Times (h) glucose xylose Rs up to 4.5 g/l/h in a fermentor ( E. coli) and still non optimal 21

22 %total DEINOCCOCUS IN THE REAL LIFE ASSIMILATION OF MIXES OF BIRCH WOOD XYLAN Xylan is directly hydrolized by the strains and assimilated 120 Kinetic of xylose assimilation on birch wood xylan % of the total xylan is consumed after 48h of cultivation

23 g/l g/l g/l g/l DEINOCCOCUS AS A SECOND GENERATION CHASSIS CELLULOLYSIS & ASSIMILATION OF SMALL CELLULOSE POLYMERS GLUCOSE CELLOTRIOSE Time(h) Time(h) Inoculated CELLOBIOSE CELLOTETRAOSE Control Activity on watman paper Time(h) Time(h) 23

24 FROM 1.5 TO 2G BIOETHANOL 24

25 THE DEINOL PROGRAM TOWARD 2 G BIOETHANOL PROCESSES IN DEVELOPMENT Enzymatic hydrolysis 70 C Fermentation 37 C YIELD ++ 2G bioethanol NON FOOD BIOMASS Proof of concept Pretreatment Distillation LIGHT DEINOL ONE STEP AT 45 C VERY ROBUST NO ADDITIVE Coproducts NUTRITIONAL QUALITY ++ 25

26 DEINOL AN ACCELERATED DEVELOPMENT 2011 PROCESS PREVALIDATION (EC1) 2012 PROOF OF CONCEPT (EC2) 2013 PILOT LAB (EC3) INDUSTRIAL PILOT (EC4) 2014 PRODUCTION The optimized strain degrade more than 80% of a wheat-based biomass into simple sugars Without additive, the optimized strain transforms the same substrate into bioethanol with an alcohol content of 3% Target: process validation in large fermenter (300L) and further optimization Industrial validation in operating plant Hard endosperm Soft endosperm Market production 1.5 & 2G BIOFUELS Aleurone Pericarp Germ Tip cap 26

27 THE WAY TO GREEN CHEMICALS 27

28 DEINOCHEM THE WAY TO ISOPRENOIDS The largest family of natural substances in the world More than 22,000 isoprene compounds Many industrial applications: creams, perfumes, detergents, anti-cancer agents Looking for new production standards Extraction from plants and chemical synthesis suffer from low yields and high production costs Deinove s synthetic platform is ready and increased resources are on the way DEINOCHEM program Resources dedicated to the program (2012 launch / 2013 acceleration) Promising early work from our laboratories Advanced discussions with many world leading industrials partners Research contracts by the end of

29 THE DEINOCOCCUS ADVANTAGES HIGHER RESISTANCE TO ISOPRENOÏDS METABOLIZATION TEST (MTT) IC 50 (G/L) ISOPRENE GERANIOL LINALOOL b-pinene E. coli 7 0,6 4 91,7 S. cerevisiae 8 0,8 4 22,5 D. radiodurans R1 21 No inhibition 25 22,5 D. geothermalis MX6-1E 12 7,8 5 ND D. geothermalis MX6-1C 14 No inhibition 7 ND D. geothermalis MC2-2A 14 No inhibition 7 ND D. geothermalis M36-7D 22 0,6 3 91,7 29

30 CONTACTS Emmanuel PETIOT CEO Tel : emmanuel.petiot@deinove.com Jean-Paul LEONETTI VP R&D Tel : jean-paul.leonetti@deinove.com 30

31 Sustainable Development DEINOVE, a member of IAR... A world class industrial cluster Agronomy dedicated to biobased products and Biorefinery Biobased feedstock Logistic & pre-treatment 210 members 40% SMEs 135 R&I projects Total R&I budget: > 1,2 bn Green Chemistry Lab Industrial Biobased products biofuels, chemicals, materials, ingredients M a r k e t s... Industrial Biotech Pilot plant demonstration