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Synthetic Consortium for Cellulose Hydrolysis and Ethanol Production

Synthetic Consortium for Cellulose Hydrolysis and Ethanol Production. By: David Pham Supervisors: Wilfred Chen and Shen-Long Tsai. Composition of Municipal Solid Waste. Biomass in MSW Paper 32.7% Yard Trimmings 12.8% Wood 5.6% TOTAL: 51.1% Conclusion

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Synthetic Consortium for Cellulose Hydrolysis and Ethanol Production

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

  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

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