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Advancements in Cellulosic
Ethanol fermentation
Pierre Basuyaux AIDA conference, February 22th, 2017
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Leaf in 2 minutes
© 2016 - ALL RIGHTS RESERVED
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Lesaffre
© 2016 - ALL RIGHTS RESERVED
Turnover: 1,8 billion €. More than 60% is
generated outside Europe, including over
40% on emerging markets
Presence in 40 countries: 55 production
plants, 40 sales offices
9300 employees representing more than 70
nationalities.
Glo
bal
pre
sen
ce
Str
on
g R
&D
200 researchers in Lesaffre R&D
center
Working with over 60 universities and
research centers worldwide
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Our milestones
© 2016 - ALL RIGHTS RESERVED
Launch of Ethanol
Red® dry yeast that
became the yeast of
reference for starch
to ethanol producers
Launch of Ethanol
Red® Cream and
Propaide™
Beginning of R&D
works for
2G ethanol yeast
Sadistil™ Plus
a new yeast offer
for sugar beet
ethanol producers
Acquisition of the
Xylose Isomerase
technology from
Butalco.
Launch of CelluX™
Commissioning of
Crescentino plant
with
CelluX™
Leaf
Acquisition of
Butalco
Joint
development
agreement with
DDNA
FT 858 L
yeast of
reference for
sugar cane to
ethanol
produced and sold
under licence
from Fermentec
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Cellux Yeast
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Cellux™ Timeline
© 2016 - ALL RIGHTS RESERVED
• XI Technology
(Butalco)
XI Technology
• XI + XDH
• Xylose used
in 72 hrs
CelluX™ 1 • Improved
inhibitor
tolerance
CelluX™ 2
• Xylose
consumption
in
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Xylose Conversion Technologies
© 2016 - ALL RIGHTS RESERVED
Xylose
Xylitol
Xylulose
Xylose
Xylulose
XDH
XI
NADPH
NADP+
NAD+
NADH
Xylose
Xylulose
XI
+
XDH
XR = Xylose Reductase; XDH = Xylitol Dehydrogenase XI = Xylose
Isomerase
Cofactor Dependent (XR/XDH) Cofactor Independent (XI) -
Cellux™
XR
Cellux Butalco IP
+ Industrial
ADY strain
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Cellux™ Advantages
© 2016 - ALL RIGHTS RESERVED
XI Technology is not cofactor dependent = No xylitol
formation
GMO strain with industrial strain characteristics
Hardy
High inhibitor tolerance
Fast xylose fermentation kinetics
Available in dry form since 2012
Successful demonstration on mash obtained from different
pretreatment technologies/substrates
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Strain improvement
by genome
shuffling
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Genome Shuffling
Genes involved in C5 metabolism
Strain with an interesting inhibitor tolerance
CelluX™1 = Xylose +
Hybrid with strong genetic background due to heterosys
Genome shuffling =
Mass sporulation + Random hybridization
production of a very diverse population
in terms of genomic sequence
© 2016 - ALL RIGHTS RESERVED
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C5 selection =
Growth on medium
containing xylose as sole carbon source
Keeping the C5 phenotype
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A new Lesaffre technology makes us able to keep the essential C5
genes
in each cell of the population
Only the strains with the best xylose consuming rate
survive
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Selecting the best strain
Enrichment on poor medium =
Growth on medium containing : Glucose Mineral nitrogen source
Inhibitors
The strains with the best genetic
background grow faster than the others
CelluX™ 3 = Xylose +
and resistance
© 2016 - ALL RIGHTS RESERVED
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Typical fermentation
results
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Fermentation trials
Partnership and collaborations with process developers /
customers for:
Strain performance evaluation
Propagation/fermentation process set-up
On site technical assistance
Numerous fermentation trials with materials coming from all over
the world
Basic process includes:
A short propagation pitched with the ADY @ 0.5 g/kg
A 10% propagation mash to 90% fermentation mash transfer
Industrial nutrients use (urea, phosphate, few minerals)
T control in the 30-35°C range
pH in the 5.0-5.5 range
© 2016 - ALL RIGHTS RESERVED
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Production &
commercialization
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Commercialization
Lesaffre has proven ability to produce and commercialize
GMO yeasts
CelluX™ yeasts are submitted to pre-market
approval (commercial release) or authorizations of use
(Class 1 of GMMs)
European Union: authorizations for contained use (at Member
State level and user specific)
US: MCAN granted to Lesaffre by EPA in Q1 2014
Under progress: Brazil, China, India
© 2016 - ALL RIGHTS RESERVED
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Grand opening
© 2016 - ALL RIGHTS RESERVED
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Next steps
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Perspectives
Strain improvement continues to fulfill partners needs and
improve the economics
Focus areas
Optimization of ethanol productivity
Improvement of the lignocellulosic biomass to ethanol yield
© 2016 - ALL RIGHTS RESERVED
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Perspectives
Specific arabinose transporter of the plant Arabidopsis thaliana
for the construction of pentose fermenting yeasts Novel specific
arabinose transporter from the yeast Scheffersomyces stipitis, and
uses thereof Vector with codon-optimized genes for an arabinose
metabolic pathway for arabinose conversion in yeast for ethanol
production
© 2016 - ALL RIGHTS RESERVED
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Thank you for your attention
www.lesaffreadvancedfermentations.com – [email protected]
