1. Speaker: Jeng-Chen Liu(劉政成) Student ID: P9921003
Graduate Institute of Bioresources, National Pingtung University of Science and Technology
Proposal for Ph. D. Program
Yeast expression of cellulase complex from rumen fungi for direct fermentation of cellulose to ethanol
Speaker: Jeng-Chen Liu(劉政成)
Student ID: P

2. Outline Background: fuel requirement & bioethanol
Goal: direct microbial conversion
Materials and methods: gene sources & expression
Expected results: cellulose  ethanol

3. The problems of petroleum utilization
Exhaustion
Petrolic resources
Global warming
fossil fuels
CO2+H2O

4. The cycle of biomass energy
Photosynthesis
Sugar Starch Lignocellulose Lipid
CO2+H2O
Renewable
Net emission(CO2)=0
Ethanol-gasoline
eg. E3
Biofuels eg. bioethanol

5. Bioethanol: first generation
Unauthorized cultivation
Food: sugar starch bean
Food price

6. Bioethanol: second generation
Unauthorized cultivation
Food: sugar starch bean
Food price

7. Bioethanol: second generation
Unauthorized cultivation
Lignocellulose
Food price

8. The contents of lignocellulose
2 M ton * 40% =800 K ton
(Prasad et al., 2007)
(Murphy and McCarthy, 2005)

9. Lignocellulose  ethanol
Pretreatment: removed lignin & hemicellulose
Hydrolysis: cellulose  glucose
Fermentation: glucose  ethanol

10. Pretreatment Physical methods Chemical methods Biological methods
Ethanol
Cellulose
Hemicellulose
Lignin
(Lu and Mosier, 2008)

11. Cellulose  glucose
Acidic hydrolysis Enzymatic hydrolysis

12. Conversion of cellulose to glucose
Table 1. Comparison of hydrolysis methods for cellulose hydrolysis
Acid hydrolysis Enzymatic hydrolysis？
Cellulose
Glucose
(Bungay, 2004)

13. Lignocellulose  ethanol(SHF;SSF;DMC)
Pretreatment: removed lignin & hemicellulose
Hydrolysis: cellulose  glucose
Fermentation: glucose  ethanol

14. Lignocellulose  ethanol(SHF)
Pretreatment
Hydrolysis: cellulase supplementation
Fermentation: microorganism
Cellulose  ethanol

15. Lignocellulose  ethanol(SSF)
Pretreatment
Hydrolysis: cellulase supplementation
Fermentation: microorganism
Cellulose  ethanol

16. Lignocellulose  ethanol(DMC)
Pretreatment
Hydrolysis: cellulase
Fermentation: microorganism
Cellulose
Glucose
ethanol

17. The analysis of enzyme hydrosylate and fermentation products
(Chen et al., 2003)
Cellulase complex
(劉，2010)

18. Glucose  ethanol Saccharomyces cerevisiae
ethanol producer ethanol tolerant GRAS
Zymomonas mobilis
Escherichia coli

19. Lignocellulose  ethanol(DMC)
Goal
Lignocellulose  ethanol(DMC)
Pretreatment
Hydrolysis
Fermentation: S. cerevisiae
Cellulose
DMC
Glucose
ethanol

20. Lignocellulose  ethanol(DMC)
Goal
Lignocellulose  ethanol(DMC)
Pretreatment
Hydrolysis: cellulase
Fermentation: S. cerevisiae
Cellulase complex gene from rumen fungi
Cellulose
DMC
Glucose
ethanol

21. Protocols Rumen fungi cellulase complex genes
E. coli expression system
S. cerevisiae expression system
Biochemical characterization
Cell surface display system
Promoter
Direct microbial conversion
Bioethanol

22. Lignocellulose  ethanol(DMC)
Expected results
Lignocellulose  ethanol(DMC)
Pretreatment
Hydrolysis: cellulase
Fermentation: S. cerevisiae
Cellulase complex gene from rumen fungi
Cellulose
DMC
Glucose
ethanol

23. Thanks for your attention