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Yeast expression of Yeast expression of cellulase complexcellulase complex from rumen fungi for from rumen fungi for direct fermentationdirect fermentation of of
cellulose to cellulose to ethanolethanol
Speaker: Speaker: Jeng-Chen Liu(Jeng-Chen Liu( 劉政劉政成成 ))
Student ID: P9921003Student ID: P9921003
Graduate Institute of Bioresources, National Pingtung University of Science and Technology
Proposal for Ph. D. ProgramProposal for Ph. D. Program
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OutlineOutline
BackgroundBackground: : fuel requirement & bioethanolfuel requirement & bioethanol GoalGoal: : direct microbial conversiondirect microbial conversion Materials and methodsMaterials and methods: : gene sources & expressiongene sources & expression Expected resultsExpected results: : cellulose cellulose ethanol ethanol
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Petrolic resources
The problems of petroleum utilizationThe problems of petroleum utilization
Exhaustion
Global warming
CO2+H2Ofossil fuelsfossil fuels
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The cycle of biomass energyThe cycle of biomass energy
CO2+H2O
Photosynthesis
SugarStarchLignocellulose Lipid
Biofuelseg. bioethanol
RenewableRenewable Net emission(CONet emission(CO22)=0)=0
Ethanol-gasoline
eg. E3
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Bioethanol: first generationBioethanol: first generation
Food: sugar starch bean
Food price
Unauthorized cultivation
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Bioethanol: second generationBioethanol: second generation
Food: sugar starch bean
Food price
Unauthorized cultivation
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Bioethanol: second generationBioethanol: second generation
Food price
Unauthorized cultivation
Lignocellulose
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The contents of lignocelluloseThe contents of lignocellulose
(Murphy and McCarthy, 2005)(Prasad et al., 2007)
2 M ton * 40%=800 K ton
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Lignocellulose Lignocellulose ethanol ethanol
Pretreatment:Pretreatment: removed lignin & hemicelluloseremoved lignin & hemicellulose
Hydrolysis:Hydrolysis: cellulose cellulose glucose glucose
Fermentation:Fermentation: glucose glucose ethanol ethanol
1010
PretreatmentPretreatment
(Lu and Mosier, 2008)
Physical methodsChemical methodsBiological methods
Lignin
Hemicellulose
Cellulose
Ethanol
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Cellulose Cellulose glucose glucose
Acidic hydrolysis Enzymatic hydrolysis
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Table 1. Comparison of hydrolysis methods for cellulose hydrolysis
(Bungay, 2004)
Conversion of cellulose to glucose
Cellulose Glucose
Acid hydrolysisEnzymatic hydrolysis？
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Lignocellulose Lignocellulose ethanol(SHF;SSF;DMC) ethanol(SHF;SSF;DMC)
Pretreatment:Pretreatment: removed lignin & hemicelluloseremoved lignin & hemicellulose
Hydrolysis:Hydrolysis: cellulose cellulose glucose glucose
Fermentation:Fermentation: glucose glucose ethanol ethanol
1414
Lignocellulose Lignocellulose ethanol(SHF) ethanol(SHF)
PretreatmentPretreatment
Hydrolysis: cellulase supplementationHydrolysis: cellulase supplementation
Fermentation: microorganismFermentation: microorganism
Cellulose ethanol
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Lignocellulose Lignocellulose ethanol(SSF) ethanol(SSF)
PretreatmentPretreatment
Hydrolysis: cellulase supplementationHydrolysis: cellulase supplementation
Fermentation: microorganismFermentation: microorganism
Cellulose ethanol
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Lignocellulose Lignocellulose ethanol(DMC) ethanol(DMC)
PretreatmentPretreatment
Hydrolysis: cellulaseHydrolysis: cellulase
Fermentation: microorganismFermentation: microorganismGlucose
Cellulose
ethanol
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The analysis of enzyme hydrosylate The analysis of enzyme hydrosylate and fermentation productsand fermentation products
(劉， 2010)
(Chen et al., 2003)
Cellulase complex
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Glucose Glucose ethanol ethanol
Saccharomyces cerevisiaeSaccharomyces cerevisiae
ethanol producerethanol producerethanol tolerant ethanol tolerant GRASGRAS
Zymomonas mobilisZymomonas mobilis
Escherichia coliEscherichia coli
1919
Lignocellulose Lignocellulose ethanol(DMC) ethanol(DMC) PretreatmentPretreatment
HydrolysisHydrolysis
Fermentation:Fermentation: S. cerevisiaeS. cerevisiae
DMCDMC
Glucose
Cellulose
ethanol
GoalGoal
2020
Lignocellulose Lignocellulose ethanol(DMC) ethanol(DMC) PretreatmentPretreatment
Hydrolysis: cellulaseHydrolysis: cellulase
Fermentation:Fermentation: S. cerevisiaeS. cerevisiae
DMCDMC
Glucose
Cellulose
ethanol
GoalGoal
Cellulase complex gene from rumen fungi
2121
ProtocolsProtocols Rumen fungi
cellulase complex genescellulase complex genes
E. coli expression system
S. cerevisiae expression system
Biochemical characterization
Promoter Cell surface display system
Direct microbial conversion
Bioethanol
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Lignocellulose Lignocellulose ethanol(DMC) ethanol(DMC) PretreatmentPretreatment
Hydrolysis: cellulaseHydrolysis: cellulase
Fermentation:Fermentation: S. cerevisiaeS. cerevisiae
DMCDMC
Glucose
Cellulose
ethanol
Cellulase complex gene from rumen fungi
Expected resultsExpected results
2323
Thanks for Thanks for your attentionyour attention
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