Dynamic Regulation and Metabolic Engineering of Total Biodiesel Production in E. coli
Poster presentation at Clemson University 10th Annual Focus on Creative Inquiry Forum, Clemson, SC.
Abstract
The purpose of the project is to engineer a strain of E. coli that produces fatty acid ethyl esters more efficiently. Esters are used in numerous industrial and commercial products including fuel, lubricants, fragrances, and flavorings. Currently there is an imbalance in the ester producing pathway of E. coli that creates an abundance of alcohol in relation to fatty acid, preventing optimal efficiency. As of now we have replaced pyruvate decarboxylase and acyl transferase genes using techniques such as polymerase chain reaction, sequence and ligation independent cloning, and electroporation. The next step of our project is to confirm increased production rates via growth studies and gas chromatography to measure the amounts of fatty acid ethyl ester produced. In the future we hope to experiment with atfA, an acyl transferase gene engineered to have higher solubility and thus more enzymatic activity. Through this we hope to achieve not only increased fatty acid ethyl ester production, but also to identify the effect of varying substrates on the type of ester produced.