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

Nonrenewable Energy Consumption Gas price increasing Food price increasing Bio-fuel is the solution Food VS Fuel Lignocellulose: nature’s carbon mine We need right tool for the job!

Anaerobic gut fungi

Brighter Future

The Big Picture

Anaerobic Gut Fungi Enzyme: Cellulase

Assemble

Cellulosome Complex Enzyme Dockerin Cohesin Scaffoldin Dockerin & Cohesin interaction Binding partner?

Research Goal:

Decipher Fungal Cellulosome Architecture by Identifying Dockerin Binding Partner

Brighter Future

The Project Procedure

Cel6A

Polymerase Chain Reaction

Denaturation Annealing Elongation DNA Polymerase

Experimental Method

Cel6A

Polymerase Chain Reaction

The Project Procedure

Protein Gel Electrophoresis

• +

25 20 15 10 Unit: Kilodalton | | 1.7×10−24 Kg Protein

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

• Use a different E.Coli strain -Tuner
• Replace the rare codons
• Try E.Coli strain that contain both features of Origami and Tuner

Time line

• Tuner is capable of recognizing rare code
• Dockerin proteins may not be folded properly

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