PRESENTATION BIO WORLD CONGRESS – 19 JUNE 2013

COMPANY SNAPSHOT

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.

DEINOVE, THE DEINOCOCCUS WAY

2

DEINOVE

KEY DATES

3

2009 2007 2008

Inception, co-founders Pr Miroslav RADMAN Dr Philippe POULETTY

2013 2006 2010

DEINOL

EC1

2011 2012

OSEO funding (€ 1.57 m)

Jacques BITON CEO

IPO (€ 12 m) Partnership with TEREOS Partnership with VTT OSEO funding (€ 1.4 m)

OSEO funding (€6 m)

Deinobank (6 000 strains) Initiation of DEINOL program

Deinobank (25 strains)

Proprietary technological platform Initiation of DEINOBIOTICS program

Initiation of DEINOCHEM program

DEINOL EC2 (POC) THANAPLAST™ program

DEINOBIOTICS SAS

OSEO funding (€ 1.15 m) Market Cap: € 53 m

Emmanuel PETIOT CEO

2 5 31 Staff

DEINOCHEM

DEINOVE

STRATEGY FROM 2012 to 2013

4

DEINOL…and beyond

Second generation biofuels

Plastics Fine chemicals

Sweeteners Fragrances Flavors

Detergents =

Other programs

DEINOBIOTICS THANAPLAST™

Antibiotics

Plastic remediation

DEINOCOCCUS: A NEW CHASSIS FOR FERMENTATION

5

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

DEINOCOCCUS: A NEW CHASSIS FOR 2nd GENERATION BIOFUELS

A UNIQUE SUM OF BENEFITS

THE GENUS DEINOCOCCUS

A NEW SCIENTIFIC AND TECHNOLOGICAL WAY

7

• The largest collection of Deinococci in the world (6 000 strains)

• A proprietary technological platform (16 patents) with validation through a POC (DEINOL)

Num

ber

of public

ations

200 000

100 000

300 000

Microorganisms

NUMBER OF PUBLICATIONS PER GENUS

Num

ber

of pate

nts

4 000

0

8 000

Microorganisms

NUMBER OF PATENTS PER GENUS

Escherichia coli

Saccharomyces cerevisiae

Deinococcus

Deinococcus geothermalis

The world pioneer of deinotechnologies

OUR PROPRIETARY TECHNOLOGICAL PLATFORM

THE BEST OF GENETIC ENGINEERING AND BACTERIOLOGY

8

ARCHIVING

Library of cryopreserved

strains

Physiology

High-throughput growth assays

Metabolomic

Functional tests Gene expression

measurement

Microbiology

Genomic

PCR Sequencing & Annotation

DEINOBANK (6 000 clones)

Genetic & Metabolic

Engineering

Gene insertion Gene deletion

Sequence optimization

LAB PILOT

INDUSTRIAL PILOT

PRODUCTION

COLLECTION SELECTION OPTIMISATION SCALE UP

LEAD DEINOCOCCUS CANDIDAT

S1 S2 S3 POC

Fermentation engineering

Optimization of culture &

fermentation processes

Industrial engineering

Lab Assays Test &

Integration

Samples collection in

extreme environment

THE DEINOVE TEAM

A YOUNG INNOVATIVE COMPANY

9

A BACTERIAL LIBRARY OF 6 000 STRAINS

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

MANY UNSUAL STRAINS DUE TO SELECTION BY IRRADIATION

10

INHIBITORS RESISTANCE

11

THE DEINOCOCCUS ADVANTAGES

Metabolization test (MTT)

IC50 (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

THE DEINOCOCCUS ADVANTAGES

A GENETIC MOSAIC WITH UNUSUAL ROBUSTNESS…

12

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

1 2 3 1 2 3

ETOH OPERON

1 Recombinant D. geothermalis 2 Recombinant D. geothermalis after serial dilution 3 Wild type D. geothermalis,

BIOMASS OPERON

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)

THE DEINOCOCCUS ADVANTAGES

… NOW ACCESSIBLE TO DIRECTED EVOLUTION

13

-20

0

20

40

60

80

100

120

0 2 5 10 15 20 25 40

OD

57

0 N

M (

% C

ON

TRO

L)

% ÉTHANOL

E. coli

S. Cerevisiae

D. radiodurans R1

Recombinant D. geothermalis

Evolved recombinant D. geothermalis

0 2 5 10 15 20 25 40

% ETHANOL

DEINOVE’S PLATFORM

14

PLATFORM

STRAIN CONSTRUCTION PLATFORM

15

Library of standard

elements: promotors

RBS…

Omic databases

genomic,

transcriptomic,

proteomic…

A large strain library

6000 strains, 1800

Deinococcus

0

1000

2000

3000

4000

5000

6000

S1 2011 S2 2011 S1 2012 S2 2012

Generated strains A robotic platform

Dedicated software Robotic cloning Deinococcus allows fast

strain construction

Rémi Bernard

Molecular Biology Manager

PLATFORM

FERMENTATION METABOLISM ANALYTIC

16

Fermentation & Metabolism

Multi parallel

fermentors:

1L, 2 L, 5L,

20L.

Enzymology,

fluxomic,

metabolomic

Analytical Platform

MicrOTOF-QII Bruker

(intra and extracellular

metabolites)

Dionex ICS (sugars, acids) Gaz

chromatorgraphy

variant

DEINOCOCCUS ENZYMES, BIOMASS DEGRADATION, SUGAR ASSIMILATION

17

BIOMASS HYDROLYZING ENZYMES

50 STRAINS SEQUENCED, PHYLOGENY OF GLYCOSYL HYDROLASES

18

Mx6

-1B

DR

H-4

6

M1-3

H

M13-1

A

MC

3-9

A

MI3

-1C

M11-1

2B

MI3

_10

M23r-

2A

M23-2

E

M31-8

F

MC

5-1

1A

MX

4-5

F

M31-1

0

M36-8

C

M19-G

G

M26-8

A

MC

3-4

A

MC

3-4

B

MC

3-5

G

MC

2-2

C

MC

2-2

A

M36-7

0

MX

6-1

C

MX

6-1

0

MX

6-1

E

DR

H-3

7

DR

H-3

8

DS

M11300

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

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(68)

3.2.1.23

3.2.1.22

3.2.1.21

23.2.1.97

3.2.1.8

3.2.1.37

3.2.1.4

3.2.1.2152)

3.2.1.40

3.2.1

GH13

GH13

GH13

GH13

GH13

GH13

GH13

GH13

GH77

GH13

GH15

GH13

GH13

GH42

GH36

GH1

GH101

GH8

GH52

GH5

GH3

GH78

GH3

BIOMASS HYDROLYZING ENZYMES

DEINOCOCCUS ENDOCELLULASE

19

Closest relative: Clostridium

cellulolyticum GH5 b1,4-glucanase

Clostridium cellulolyticum (65% identity)

1 125 250 375 500 625 750 806

QC

Cellulase

Cellulase superfamily

SH

SpF CBM 1728 CBM 1728

Control (no bacteria)

Deinococcus

AZO-CMC (cellulose 0.5%)

BIOMASS HYDROLYZING ENZYMES

GH10 XYLANASE

20

Glycoside hydrolase family 10, closest relative Clostridium thermocellum (45% identity)

1 50 100 150 200 250 300 344

QC

Glyco hydro 10

Glyco hydro 10 superfamily

SH

SpF

AZO-Xylan (0.5%)

DEINOCCOCUS IN THE REAL LIFE

ASSIMILATION OF MIXES OF GLUCOSE & XYLOSE

21

0

20

0.2 0

40

60

80

100

120

0.4 0.6 0.8 1 1.2

% OF RS MAX

Ratio glucose/xylose g/g

Rs up to 4.5 g/l/h in a fermentor (≈ E. coli) and still non optimal

0

2

4

6

8

10

12

0 10 20 30 40 50 60 70 80

Fre

e s

ug

ars

(g

/l)

Times (h)

WHEAT BRAN, shake flask

glucose xylose

22

• Xylan is directly hydrolized by the strains and assimilated

0

20

40

60

80

100

120

0 50 100 150 200

%to

tal

Kinetic of xylose assimilation on birch wood xylan

80% of the total xylan is

consumed after 48h of

cultivation

DEINOCCOCUS IN THE REAL LIFE

ASSIMILATION OF MIXES OF BIRCH WOOD XYLAN

DEINOCCOCUS AS A SECOND GENERATION CHASSIS

CELLULOLYSIS & ASSIMILATION OF SMALL CELLULOSE POLYMERS

23

0

2

10 0

4

6

8

10

12

14 16

20 30 40 50 60 70 80

GLUCOSE

Time(h)

g/l

0

2

10 0

4

6

8

10

12

14 16

20 30 40 50 60 70 80

CELLOBIOSE

Time(h)

g/l

0

2

10 0

4

6

8

10

12

14 16

20 30 40 50 60 70 80

CELLOTRIOSE

Time(h)

g/l

0

2

10 0

4

6

8

10

12

14 16

20 30 40 50 60 70 80

CELLOTETRAOSE

Time(h)

g/l

Activity on watman paper

Inoculated

Control

FROM 1.5 TO 2G BIOETHANOL

24

THE DEINOL PROGRAM

TOWARD 2 G BIOETHANOL

25

PROCESSES IN

DEVELOPMENT

DEINOL

NON FOOD BIOMASS

Distillation

2G bioethanol

Coproducts

Pretreatment

Enzymatic hydrolysis 70°C

Fermentation 37°C

NUTRITIONAL QUALITY ++

LIGHT

YIELD ++

ONE STEP AT 45°C VERY ROBUST NO ADDITIVE

Proof of

concept

DEINOL

AN ACCELERATED DEVELOPMENT

26

2011 PROCESS

PREVALIDATION (EC1)

The optimized strain degrade

more than 80% of a wheat-based biomass into simple sugars

2012 PROOF OF CONCEPT

(EC2)

Target: process validation in large fermenter (300L) and further

optimization

2013 PILOT LAB

(EC3)

Without additive, the optimized

strain transforms the same substrate

into bioethanol with an alcohol content of 3%

2013-2014 INDUSTRIAL PILOT

(EC4)

2014 PRODUCTION

Industrial validation in operating plant

Market production

Soft endosperm Hard endosperm

Germ

Aleurone

Pericarp

Tip cap

1.5 & 2G BIOFUELS

THE WAY TO GREEN CHEMICALS

27

DEINOCHEM

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 2013

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

THE WAY TO ISOPRENOIDS

28

THE DEINOCOCCUS ADVANTAGES

HIGHER RESISTANCE TO ISOPRENOÏDS

29

IC50 (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

METABOLIZATION TEST (MTT)

CONTACTS

30

Emmanuel PETIOT CEO Tel : + 33 1 42 03 27 37 emmanuel.petiot@deinove.com

Jean-Paul LEONETTI VP R&D Tel : + 33 1 42 03 27 37 jean-paul.leonetti@deinove.com

………………………………………………………………………………………….........

………………………………………………………………………………………….........

DEINOVE, a member of IAR

• 210 members

• 40% SMEs

• 135 R&I projects

• Total R&I budget: > 1,2 bn €

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