BIOMASS TO FUEL:PROBLEMS AND SOLUTIONS

Dr. Steve Hutcheson, PhDDept. Cell Biology and Molecular Genetics

BIOFUEL CARBON CYCLE

CO2

1YEAR

CONVERSION

BIOMASSBIOETHANOLBIODIESELBIOBUTANOLBIOMETHANE

CORN-BASED ETHANOL

Grain milled to release corn starchGrain milled to release corn starchGrain milled to release corn starch

EnzymesEnzymesEnzymes

GlucoseGlucoseGlucose

FermentationFermentationFermentation

Distillation Distillation Distillation

© Zymetis 2007© Zymetis 2007

α 1,4 glucose +α 1,6 branchesαα 1,4 glucose +1,4 glucose +αα 1,6 branches1,6 branches

BIOETHANOLBIOETHANOLBIOETHANOL

Yield: 89 gallons/ton375 gal/acreYield: 89 gallons/tonYield: 89 gallons/ton375375 gal/acregal/acre

GLUCOSE ->2 CH3CH2OH + 2 CO2

GLUCOSE GLUCOSE -->>2 CH2 CH33CHCH22OH + 2 COOH + 2 CO22

ETHANOL PRODUCTIONIN THE USA

ETHANOLETHANOLETHANOL

~2.2 billion bushels~2.2 billion bushels~2.2 billion bushels

CURRENTCURRENTCURRENT

~5.5 billion gallons~5.5 billion gallons~5.5 billion gallons

TOTAL US CORN PRODUCTIONTOTAL US CORN TOTAL US CORN PRODUCTIONPRODUCTION

~10 billion bushels~10 billion bushels~10 billion bushels

THEORETICAL MAX YIELD: ~25 billion gallonsTHEORETICAL MAX THEORETICAL MAX YIELD: ~25 billion gallonsYIELD: ~25 billion gallons

130Facilities130130FacilitiesFacilities

© Zymetis 2007©© Zymetis 2007Zymetis 2007

FOODFOOD

17% ofFuel Use17% ofFuel Use

Low EGain

Low EGain

ETHANOL FROM GRAIN IS WASTEFUL

Grain milled to release corn starchGrain milled to release corn starchGrain milled to release corn starch

EnzymesEnzymesEnzymes

GlucoseGlucoseGlucose

FermentationFermentationFermentation

Distillation Distillation Distillation

© Zymetis 2007© Zymetis 2007

α 1,4 glucose +α 1,6 branchesαα 1,4 glucose +1,4 glucose +αα 1,6 branches1,6 branches

BIOETHANOLBIOETHANOLBIOETHANOL

Yield: 89 gallons/tonProduction cost: $~1.20/gallonYield: 89 gallons/tonYield: 89 gallons/tonProduction cost: $~1.20/gallonProduction cost: $~1.20/gallon

GLUCOSE ->2 CH3CH2OH + 2 CO2

GLUCOSE GLUCOSE -->>2 CH2 CH33CHCH22OH + 2 COOH + 2 CO22

>80% biomass>80% biomass>80% biomass WASTES(REST OF PLANT)

WASTESWASTES(REST OF PLANT)(REST OF PLANT)

POTENTIAL CELLULOSIC ETHANOL PRODUCTION

Estimated >75 billion gallons/year renewable

resource

Estimated >75 billion Estimated >75 billion gallons/year renewable gallons/year renewable

resourceresource

© Zymetis 2007

Corn Corn stoverstover

300 gal/acre300 gal/acre300 gal/acre

SwitchgrassSwitchgrass

800 gal/acre800 gal/acre

Wheat strawWheat straw

100-150 gal/acre100100--150 gal/acre150 gal/acre

ForestryForestrybyproductsbyproducts

1000 gal/acre1000 gal/acre

BIOMASS CONVERSION METHODS

ENZYME-MEDIATED

GASIFICATION

��STARCHα 1,4 glucose +α 1,6 branches

����STARCHSTARCHαα 1,4 glucose +1,4 glucose +αα 1,6 branches1,6 branches

CORN IS CHEMICALLY SIMPLE

http://micro.magnet.fsu.edu/cells/plants/images/

http://www.emc.maricopa.edu/faculty/farabee/BIOBK/cellulose.gif

BIOMASS ISCHEMICALLYCOMPLEX

CELLULOSE

ββ 1,4 glucose1,4 glucose

CELL WALL POLYSACCHARIDES

CELLULOSE

LIGNIN

BARRIERS TO PROCESSING

FERMENTABLEC6 & C5 SUGARS

HEMICELLULOSE & PECTINS

X X X X X X X X X X X X X X

XG

al

XAA

A AAX A

A

Gal

A

Guc Ac

Gal

Gla

R

Gla

Gal

A

Gal

A

R

Gla

R

Gla

R

Gla

R

Gla

R

Gla

R

Gla

R

A

A

A

A

A

A

A AA

A

Fuc

Gal

Gal

Gal

Gal

Gal

Gal

LigninLignin

http://micro.magnet.fsu.edu/cells/plants/images/

Hemicellulose Pectin

ENZYMIC BIOETHANOL PRODUCTION

Pretreatment

Processed Material

C6 Sugars

Fermentation

Distillation SIEVE© Zymetis 2007

MillingLignin & hemicellulose

extraction (hot H2SO4)Steam explosion

BIOETHANOL

Lignin(Fuel)

C5 sugars

FEEDSTOCKCELLULOSE

LIGNIN

BARRIERS TO PROCESSING

FERMENTABLEC6 & C5 SUGARS

HEMICELLULOSE & PECTINS

EnzymesEnzymesEnzymes

ETHANOL PRODUCTION

6 C sugars6 C sugars6 C sugars

>34 moles

68 moles

One gallon

11 kg

9 kg

10 kg

$2.50

$1.50

$0.60

$0.44

TARGETENZYME < $0.25COST

ECONOMICAL SOURCES OF ENZYMES?

SOURCES OF CELLULOLYTIC ORGANISMS

Saccharophagus degradans 2-40

• Isolated from decaying marsh grass

• Gamma-proteobacterium (Pseudoalteromonas group)

• Aerobic, marine bacterium

• Unusual degradative abilitiesAndrykovich & Marx, 1988

CPs Degraded

Higher Plant Polysaccharides

Cellulose Xylan

Pectin Starch

Algal Polysaccharides

Agar Pullulan

Laminarin Fucoidan

Biofilm Components

Alginate

Exoskeleton and Fungal Components

Chitin

GENOMICS

• 5.06 MB chromosome

• 4009 gene models

• 128 genes with GH domains33 genes with PL domains

• Accessory genes

A conservative estimate: ~10% of the genome is dedicated to polysaccharide depolymerization, transport, and metabolism

Courtesy of L. Hauser, F. Larimer, and P. Richardson

6 C sugars6 C sugars6 C sugars

5 C sugars5 C sugars5 C sugars

CELL WALL PROCESSING ENZYMES PREDICTED FROM THE GENOME SEQUENCE

©© Zymetis 2007Zymetis 2007

ONESTOPSHOP!!!

ONEONESTOPSTOPSHOP!!!SHOP!!!

CELLULOSE

LIGNIN

HEMICELLULOSE & PECTINS

• Xylanases (7)• Xylosidases (8)• Arabinofuranosidases (11)• Acetylxylan esterases (2)• Mannases (5)

•• Xylanases Xylanases (7)(7)•• Xylosidases Xylosidases (8)(8)•• Arabinofuranosidases Arabinofuranosidases (11)(11)•• Acetylxylan esterases Acetylxylan esterases (2)(2)•• Mannases Mannases (5)(5)

• Pectate lyases (14)• Pectin methyl esterases(2)• Arabinases (10)• Galactosidases (2)• Glucuronidases (2)

•• Pectate lyasesPectate lyases (14)(14)•• Pectin methyl Pectin methyl esterasesesterases(2)(2)•• Arabinases Arabinases (10)(10)•• Galactosidases Galactosidases (2)(2)•• Glucuronidases Glucuronidases (2)(2)

• Endoglucanases (12)• Cellobiohydrolase (1)• Cellodextrinases (2)• Glucanases (3)• CBM proteins (>10)

•• Endoglucanases Endoglucanases (12)(12)•• Cellobiohydrolase Cellobiohydrolase (1)(1)•• Cellodextrinases Cellodextrinases (2)(2)•• Glucanases Glucanases (3)(3)•• CBM proteins (>10)CBM proteins (>10)

Ligninases (3-5)Ligninases Ligninases (3(3--5)5)

DIGESTION OF PLANT MATERIAL DURING CULTURE WITH S. DEGRADANS

NEWSPRINT

CORN LEAVES

© Zymetis 2007

ENZYMES FOR FUELSENZYMES FORENZYMES FOR FUELSFUELS

(Zy - met - is)

BIOMASS CONVERSION METHODS

ENZYME-MEDIATED

GASIFICATION

SYNTHETIC “BIOETHANOL”

HIGHTEMP

GASIFIER

(1800 F)

SYNGASCATALYTIC CONVERSION

BIOFUEL CARBON CYCLE

CO2

1YEAR

CONVERSION

BIOMASSBIOETHANOLBIODIESELBIOBUTANOLBIOMETHANE

BIODIESEL

LyeMethanol

OIL SEED

4% original biomass

OIL

POTENTIAL BIODIESEL YIELDS

16,700 sq mi7000Algae

190,000 sq mi635Oil palm

1 million sq mi0.18113Peanuts

2.5 million sq mi548Soybean

Total(billion gal)

Gal / acre

Land area50% need

YieldCrop

BIOBUTAN0L PRODUCTION

Pretreatment

Processed Material

C6 Sugars

Fermentation

Distillation SIEVE© Zymetis 2007

MillingLignin & hemicellulose

extraction (hot H2SO4)Steam explosion

Lignin(Fuel)

C5 sugars

FEEDSTOCKCELLULOSE

LIGNIN

BARRIERS TO PROCESSING

FERMENTABLEC6 & C5 SUGARS

HEMICELLULOSE & PECTINS

EnzymesEnzymesEnzymes

butanol

ANAEROBIC DIGESTORS

BIOMASS

METHANE

BACTERIA

BIOFUEL CARBON CYCLE

CO2

1YEAR

CONVERSION

BIOMASSBIOETHANOLBIODIESELBIOBUTANOLBIOMETHANE

ACKNOWLEDGEMENTS

• Ron Weiner

• Loren Hauser, Frank Larimer & Paul Ricardson, DOE-JGI

• Bernie Henrissat & Pedro Coutinho (CAZY)

• Harry Gilbert & Al Boraston

• Ilya Borovok, Ed Bayer and Raphael Lamed

• D. Wilson

• Larry Taylor• Haitao Zhang• Brian Watson• A. Garrett Longmire• Jonathan Motley• Nate Ekborg• Mike Howard• Yan Shi