for Lignocellulose Breakdown Jiehao Chen Chemical Engineering East - PowerPoint PPT Presentation

Engineering Anaerobic Gut Fungi for Lignocellulose Breakdown Jiehao Chen Chemical Engineering East Los Angeles College Faculty Advisor: Dr. Michelle O'Malley Mentor: Charles Haitjema Funding Source: The U.S. Army

The Big Picture Nonrenewable Energy Consumption We need right tool for the job! Gas price increasing Anaerobic gut fungi Food price increasing Bio-fuel is the solution Food VS Fuel Brighter Future Lignocellulose: nature’s carbon mine

Assemble Cellulosome Anaerobic Gut Fungi Enzyme: Cellulase Complex Enzyme Dockerin Brighter Cohesin Future Scaffoldin Dockerin & Cohesin interaction Research Goal: Decipher Fungal Cellulosome Binding partner? Architecture by Identifying Dockerin Binding Partner

The Project Procedure Cel6A Polymerase Chain Reaction

Experimental Method DNA Polymerase Denaturation Annealing Elongation

The Project Procedure Cel6A Polymerase Chain Reaction

Protein Gel Electrophoresis Unit: Protein Kilodalton | | 1.7 × 10 −24 Kg - 25 20 15 + 10

Conclusion  Origami is not the best host for Cel6A dockerin protein expression • Cel6a has rare codons • In origami, transport RNA for rare codons are not efficient Dockerin protein wasn’t expressed very well I’m lonely!

What the future holds o Use a different E.Coli strain -Tuner • Tuner is capable of recognizing rare code • Dockerin proteins may not be folded properly o Replace the rare codons Time line o Try E.Coli strain that contain both features of Origami and Tuner

Acknowledgement • The O’Malley Lab  PI: Dr. Michelle O’Malley  Mentor: Dr. Charles Haitjema  Co-worker: Kane Nania • INSET staff • Friends • ICB • U.S. Army

Kanamycin