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May 2012
WE TURN DNA INTO PRODUCTS
DYADIC INTERNATIONAL, INC.
(Symbol: DYAI)
35TH ANNIVERSARY
2
Safe harbor statement
Certain statements contained in this presentation are forward-looking
statements. These forward-looking statements involve risks and uncertainties
that could cause Dyadic’s actual results, performance or achievements to be
materially different from any future results, performance or achievements
expressed or implied by such forward-looking statements. Except as required
by law, Dyadic expressly disclaims any intent or obligation to update any
forward-looking statements.
From Blue Jeans to Genes
3
Founded as Supplier
of Pumice Stones For
Blue Jeans
Dyadic’s C1
Expression System
1979 1992 Today
Search for Fungi
in Russia
Dyadic business overview
4
Cellulosic Sugars Industrial Enzymes Biopharmacuticals
End
mark
ets
Mark
et
siz
e
Part
ners
&
licensees
Biofuels
Bio-based chemicals
Animal feed
Food & Beverage
Starch & Alcohol
Pulp & Paper
Textiles
Vaccines
Antibodies (e.g. mAb)
Therapeutic proteins
Biocatalysts for API
$5+ billion today
$5+ billion new
enzymes in
next decade
$90+ billion today
Major
animal
health &
nutrition
company
Market
leading
food and
enzyme
company
Leading
animal
health &
nutrition
company
Leading
pharmaceutical
company
Pro
ducts
Alternafuel CMAX5
On-site licensing model
Enzyme sales to 35+
countriesPreclinical drug
development
(ABGOY)
(CDSX)
(RDS.A)
(BASFY)
(SNY)
THE C1 EXPRESSION
SYSTEM
5
Based on the Myceliophthora thermophila fungus, a
soil-borne saprophyte
Development began in the early 1990’s through a
fortuitous UV-induced mutation and continuously
bioengineered for over 20 years
White Strain developed in 2010’s to express pure proteins
A robust and versatile platform for gene discovery,
expression and the production of enzymes and other
proteins
Enables new product introduction with less time, cost
and risk by addressing critical bottlenecks of protein
discovery, development scale-up and commercialization
6
M. thermophila expression system
C1 fungus
Wild Type and new strains
WT
LC
HC
“Expression systems are not everything, but
everything is nothing without an expression
system”
DYADIC CONFIDENTIAL
C1 expression system advantages
7
Track record of producing at industrial scale
Identified over 500 potentially useful enzymes in genome, of which over
100 already been expressed
Many enzymes active in broad pH and temperature range
Excellent safety record and has Generally Recognized as Safe (GRAS)
status acknowledged by the FDA
Platform
technology
Extensive molecular toolbox with genome sequenced and annotated
Programmable to produce tailored “pure” enzymes and mixtures
Effective homologous and heterologous gene expression
Serves as both a research and production host
Cost
effective
product
development
Freedom to
operate
Dyadic is the sole owner of the C1 expression system and has broad
patent claims
C1 has two commercial strain platforms – LC and HC
8
Over-
HC LC expressed
LC
HC vs. LC
End
Market 1
Enzyme 2
(thermostable)
(12 g/L)
Enzyme 3
(thermostable)
(15 g/L)
LC-1 LC-2 LC-3
End
Market 2
LC-4
White Strain (LC) product development
Enzyme 1
(20 g/L)
The White Strain expresses dedicated enzyme mixture
Composition can be easily adapted for various
applications
High production levels and large scale fermentations
HC has ability to
effectively degrade or
modify a wide range of
lignocellulosic feedstocks
Enzyme productivity improvements by LC generation
C1-cellulase accepted by FDA on
September 29, 2009
GRAS Notification letter is a public
statement by FDA acknowledging Dyadic’s
safety determination for the intended uses
of C1
GRAS Notification letters are broadly
recognized in the food and consumer
products industries as the safety standard
C1 strain non-toxic
Pathogenicity and toxigenicity data:
strain is non-infectious and no
known toxins are produced
Peer-reviewed scientific literature
have confirmed — no known
pathogencity
No mycotoxins found
C1 enzyme testing
In vivo feeding trails:
14 day dose study in rats
13 week subchronic rat study
Genotoxicity testing:
AMES bacterial mutagenesis
Chromosomal aberration test
Genetic mutation test
No adverse effects observed
No foreign DNA
Safety confirmed
C1 has an excellent safety profile
Generally Recognized as Safe (GRAS) status
acknowledged by the FDA
10
C1 producing enzymes at industrial scale since 1996
Major C1 enzyme manufacturing sites
Year production began Manufacturing site Company Country Fermenter size
1996 FermPro/Martek (now DSM) Dyadic USA 150,000L
2000 Polfa Tarchomin Dyadic Poland 50,000L
2009 EnMex Dyadic Mexico 60,000L
2011 Antibioticos Abengoa Spain 50,000L
2011 Fermic Codexis Mexico 25,000L
2011 Iogen Codexis Canada 150,000L
2014 ADM Abengoa USA 500,000L
11
Proven industrial scale for nearly two decades
Scientific media recognition
“In 2011, a consortium of several worldwide renown organizations suggested that the use of thermostable
enzymes offers economic advantages in the production of many chemicals and biomass-based fuels.
Unfortunately, most of the commercially available enzymes which are produced by fungi like Trichoderma
and Aspergillus are not sufficiently thermostable...The reported findings confirm previous results
obtained by enzyme manufacturer Dyadic, which was the first company to successfully develop a thermophilic
fungal production platform.”
Biofuels International, March 2012
“The best studied and most widely used cellulases and hemicellulases are produced by Trichoderma,
Aspergillus...and they are most effective over a temperature range from 40°C to ~50°C. At these temperatures,
complete saccharification of biomass polysaccharides...requires long reaction times...One way to overcome these
obstacles is to raise the reaction temperature...However, implementing higher reaction temperatures requires
the deployment of enzymes that are more thermostable...Thermostable enzymes and thermophilic cell
factories may afford economic advantages in the production of many chemicals and biomass-based fuels.
Nature Biotechnology, October 2011
“The search for novel and/or improved industrial enzymes and enzyme production systems is intensifying as
market demand increases. One such new system was developed based on a recently discovered fungal isolate,
C1...The filamentous fungus C1 was developed into a mature technology and protein-production platform. C1’s
inherent richness of genes encoding industrially relevant enzymes and its high-producing characteristics
have been a proven starting point for the development of different C1 strains producing enzymes and
enzyme mixtures.”
Industrial Biotechnology, June 2011
Numerous publications have validated Dyadic’s C1 technology
12
CELLULOSIC BIOFUELS AND
BIO-BASED CHEMICALS
13
14
Commercial production of cellulosic ethanol in Q3 2014
Abengoa’s 25 million gallon plant in Hugoton, Kansas came online in 2014 and is
using Dyadic C1 enzyme technology
C1 is one of only three leading enzymatic platforms
15
Improved bioethanolenzyme mixtures
0 20 40 60 80 100
2-fold reduction
4-fold reduction
8-fold reduction
C1-G3C1-G5
CMAX CMAX 2
CMAX 3
CMAX 4
Time (months)
CMAX 5
♦
Re
lative
en
zym
e lo
ad
ing
16
Over 5-fold decrease in CMAX enzyme use/costs in 5 years
Enzyme dose reduction curve (glucose release)
17
Dyadic C1 advantages for lignocellulosic enzymes
Robust (high temperature and pH)
Highly scalable production
Versatile genetic make-up
On-site licensing model
18
Energy cropSugar Cane Corn Stover Wood Pulp Corn FiberWheat Straw DDG’sPaper pulp
Different
Pretreatments
EG’s XylanasesβGAccessory
EnzymesCBH’s
Dedicated
CMAX enzyme
mixes
High yield of
fermentable
sugars
C1 enzymes can be tailored for a variety of feedstock
Higher temperatures
19
Re
lative
activity (
%)
Temperature (°C)
05
101520253035404550556065707580859095
100
35 50 55 60 65 70
re
lativ
e a
ctiv
ity
(%
)
temperature (°C)
Relative activity after 72 hours of saccharification
G7 B7
G7 D9
Cellic Ctec 2
CMAX
Competitor
(Trichoderma-
based)
0
10
20
30
40
50
60
70
80
90
100
35 50 55 60 65 70
05
101520253035404550556065707580859095
100
35 50 55 60 65 70
re
lativ
e a
ctiv
ity
(%
)
temperature (°C)
Relative activity after 72 hours of saccharification
G7 B7
G7 D9
Cellic Ctec 2
T= 55˚C;
pretreated corn
stover (10% dry
matter)
Ability to add enzyme in early stage after
thermo-pretreatment
Broader pH
CMAX is more robust than other enzymes
CMAX3
hours
20
0
5
10
15
20
25
30
35
50 100 150
% inhib
itio
n
glucose (g/L)
CMAX3
Competitor Gen. 3
Viscosity reduction occurs during the
first hour of saccharification
0
10
20
30
40
50
60
70
80
90
0
5
10
15
20
25
30
35
40
45
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75
Perc
enta
ge d
ecre
ase
[%]
Torq
ue [%
cont
]
Time [h] Torque [cont%] Decrease [%]
0
10
20
30
40
50
60
70
80
90
0
5
10
15
20
25
30
35
40
45
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75
Perc
enta
ge d
ecre
ase [
%]
Torq
ue [%
cont
]
Time [h] Torque [cont%] Decrease [%]
Torq
ue [
% C
ont.]
Torq
ue d
ecre
ase (
%)
Time (hours)
Glucose test: T=50°C; pretreated corn stover
(10% dry matter); 72 hour fermentation
Viscosity test: Pretreated corn stover (20%
dry matter)
Lower glucose inhibition Rapid viscosity reduction
Highly scalable production
C1 has more enzyme-encoding genes than competing
systems for biofuels and bio-based chemicals
Number of lignocellulosic enzyme-encoding genes
21
Large variety of lignocellulolytic enzymes enables development of efficient dedicated enzyme
mixtures (produced by a single engineered C1-strain)
Genes encoding Number
in C1
Number in
Trichoderma
Endo-glucanases, Cellobiohydrolases, β-glucosidases/ β-xylosidases
(GH1, GH3, GH5, GH6, GH7, GH12, GH45)
~ 32 ~ 32
Cellulose binding domains (CBM1-type) ~ 46 ~ 15
Xylanases/Xylosidases (GH10, GH11, GH30,
GH43)
~ 13 ~ 10
Polysaccharide monooxygenases (GH61) ~ 26 ~ 3
Oxidoreductases (Secreted flavin-, heme-
and copper-dependent)
~ 69 ~ 24
Source: Literature and JGI database searches
Illustrative biofuel enzyme production costs
19
25
54
On-site Off-site
Shipping and distribution
Downstream processing and stabilization
Quality assurance
Utilities
Labor and overhead
Materials
Assumes 5 mg/g loading to 10% dry matter, molasses-based medium; sorbitol stabilization agent
On-site production cost advantage
22
BIOPHARMACEUTICALS
23
24
Influenza vaccine
(human)
Viral influenza
vaccine
(veterinary)
Antibodies
(diagnostic and
therapeutic)
C1 has many high-potential opportunities in biologics
Prospective game-changing platform for $90+ billion market
Biocatalysts for
API production
Therapeutic
proteins
Antiviral (HIV and
other)
Extremely high yield, industrial scale production in world’s largest
fermenters
Ability to more quickly modify cell lines to create and modify biologics
Near human glycosylation
Track record of expressing therapeutic proteins in C1
Potential to fight epidemics at global scale
Fast response to an emerging new strains of infectious disease at global scale
Origin of recent
diverse or serious
outbreaks of
emerging
infectious diseases
25
26
Key advantages throughout development cycle
Expression
Direct and single
step
transformation
Short time to
access protein
expression
Can be used for
various high-value
Rx proteins today
Already proven
for human
antibodies
Production
High production
level
Adaptable to
current reactors
Low cost of media
Short
fermentation
times
Wide range of
growth conditions
Low viscosity
Lower capex by
using smaller
facilities
Downstream
processing
Target protein
secreted into
media
Range of low
protease
production strains
Product
attributes
Favourable
glycoprofile can
be modified to
become ‘human
neutral’
Naturally
afucosylated
Proteins produced
have exhibited
excellent stability
No concern for
viruses
27
Project with Sanofi Pasteur
Vaccine using C1 expression systemTarget
Preclinical testingCurrent Phase
Expressed, purified and delivered
protein to Sanofi
Modifications to the protein are ongoing Status
Project began in 2011
Project extended and funding continues
Additional preclinical trials to follow
Timeline and
next steps
THANK YOU
28
