David B. Levin

Department of Biosystems Engineering

University of Manitoba

Winnipeg, MB

Canada

World Congress for industrial Biotechnology

June 17th – 19th, 2013

Microbial Genomics of Biocatalysts for

Consolidated Bioprocessing

Microbial Genomics of Biocatalysts for

Consolidated Bioprocessing

Slide 2

Outline

Biorefining and Consolidated Bioprocessing

Advantages of Consolidated Bioprocessing

Barriers to Consolidated Bioprocessing

Panel Speakers

BioRefining

Slide 3

Biofuels & Bioproducts from Biomass

Slide 4

BioRefining

Slide 5

Biomass Processing to Fuels & Products

BioRefining

Slide 6

The composition of lignocellulose

Cellulose is the most abundant biopolymer on earth

Cellulosic Biofuels

Slide 7

Enzymatic

Hydrolysis

To monomers

Yeast fermentation

Ethanol

+ CO2

Cellulosic Biofuels: Alternative Approach -> CBP

Slide 8

Current cellulosic biofuel technologies:

SHF: Separate Hydrolysis and

Fermentation

SSF: Simultaneous Saccharification

and Fermentation

SSFC: Simultaneous Saccharification and

CoFermentation

CBP: Consolidated BioProcessing

From Lynd et al., Microbiol Mol Biol Rev 66, 506-577

Cellulosic Biofuels:

Current vs Alternative Approach

Slide 9

Use low cost waste feedstocks

Improved energy balance

Reduced production costs

Cellulosic Biofuels: Alternative Approach -> CBP

Slide 10

Minimal

Pretreatment

Increasing access to the polymers

Simultaneous

Hydrolysis and

Fermentation

Barriers to CBP

Slide 11

NAD(P) + NAD(P)HNADPH NADP+

Formate

Fdo

Acetyl -CoA

Acetate

Fdr

Acetyl -PEthanol

CO2 H

2

NADH

NAD +

H+

AMP

+PPi

ATP

2 NADH

2 NAD +

ATP

Pi

ADP

PFO

PFL

Fd H2ase

NAD+

NADH

NAD(P)H H2ase

-16 kJ/mol -19 kJ/mol

-6 kJ/mol

-9 kJ/mol

+9 kJ/mol

-13 kJ/mol

+3 kJ/mol

+18 kJ/mol

-22 kJ/mol

PyruvateLactate

NAD + NADH

LDH

-26 kJ/mol

Cellobiose

EMP

2ATP + ADP + AMP + Pi + PPi

4ATP

2 NAD+

2 NADHFrc-1,6 -P

+

Cthe335 -343

Cthe426 -430

Cthe3003 - 3004

Cthe505

Cthe345

Cthe1035

Cthe3013 - 3024 Cthe2392

Cthe2796

Cthe3120

Cthe2430 - 2435

NFO

PTACthe1029

ACKCthe1028

ATKCthe551

Cthe423

Cthe101

Cthe394

Cthe2579

AcDH /

ADH

MalateMECthe344

OxaloacetateMDHCthe345

1) Need robust industrial biocatalysts

Limitations of strict anaerobes

Grow more slowly and to lower cell densities due to limited ATP production

“Complex” genomes

Need deeper understanding

of genome and metabolism

Slide 11

Barriers to CBP

Slide 12

2) Substrate structure influences conversion efficiency

CPB promised hydrolysis and fermentation in one step, but some pretreatment is

required

What is the minimum pretreatment required to permit access to cellulose &

hemicellulose sugars and promote efficient conversion?

Available cellulose (mg): 19.4 11.5 23.0 46.0 15.7 15.8 18.3 15.4 17.3 7.7

No pre-

treatment

Barriers to CBP

Slide 13

Efficiency of substrate conversion is a function of the degree of cellulose

crystallinity

Contribution of Amorphic Cellulose (CAC) determined by X-ray diffraction

a-cellulose

Processed wood cellulose (PWC)

Processed hemp cellulose (PHC)

Processed flax

cellulose (PFC)

Barriers to CBP

Slide 14

3) Cell/Substrate interactions are very important

Role of biofilms in substrate conversion and product synthesis?

C. Thermocellum

Panel Speakers

Slide 15

1) Richard Sparling, University of Manitoba:

Microbial Genomics for the Development of Biocatalysts for Lignocellulosic

Biorefining and Biofuels production

2) Gideon Wolfarrdt, Ryerson University:

Bacteria-substrate interaction during cellulosic conversion

3) Albrecht Laufer & Jorg Riesmeier, Direvo:

Direvo’s BluCon® high-temperature consolidated bioprocess, a substrate and

product flexible approach to low-cost biofuels and biochemicals

4) Jean-Paul Leonetti & Emmanuel Petoit, Deinove:

Development of the Deinococci C6 and C5 Consolidated Bioprocessing

Platform for Manufacturing Cellulosic Ethanol and Biobased Chemicals

page 11

Thank-you

Questions?