1. Synthetic Consortium for Cellulose Hydrolysis and Ethanol Production By: David Pham Supervisors: Wilfred Chen and Shen-Long Tsai

2. Composition of Municipal Solid Waste  Biomass in MSW Paper 32.7% Yard Trimmings 12.8% Wood 5.6%  TOTAL: 51.1%  Conclusion Over 51.1% can be hydrolyzed into ethanol Environmental Protection Agency

3. Benefits  1. Reduce overflow of Municipal Solid Waste  2. Reduce dependence on fossil fuels  3. Increase Living standards

4. Plant Biomass  Lignocellulose makes up 90% of the dry weight of plant matter  Lignocellulose average composition 45% Cellulose 30% Hemicellulose 25% Lignin

5. Cellulose

6. Benefits of a consortium  Reduced Cell Exhaustion  Environmental Resilience  Production Parameters malleable

7. Yeast Consorta to Construct

8. Cost and Efficiency of Consolidated Bioprocessing CBP costs more than three times less than current practices

9. Vector pCEL15 Figure 5. Shuttle vector between E-coli and yeast  Dockerin inserted between XhoI and BglII  EG1 inserted between EcoRI and HindIII

10. The Basics of Molecular Cloning  PCR : Replication target insert (Endoglucanase 1 (EG1) or Dockerin)  Digestion : Cutting of insert and vector (pCEL15) to provide a common conjunction  Gel Purification: Removal of digestion enzymes (Xho1, BglII, EcoR1, and HindIII)

11. The Basics of Molecular Cloning  Ligation: Fusion of insert and vector  Transformation: Delivery of plasmid into the target cell  Inoculation: Replication of the target cell  Mini-prep: Purification of plasmid from cellular components for analysis

12. Agarose Gel after Mini-Prep

13. References  EPA Municipal Solid Waste Generation, Recycling, and Disposal in the United States: Facts and Figures for 2007  www.uair.edu/botany/botimages.html  TRENDS in Biotechnology  Current Trends in Biotechnology  Microbial Biotechnology: Glazer, Nikaido 2007

14. Acknowledgements  Dr. Wilfred Chen  Shen-Long Tsai  Anjali Mulchandani  Jun Wang  BRITE Personnel  NSF