The medium-chain fatty acid (MCFAs) have a favorable safety profile and are used to treat a variety of disorders such as benefits in skin care, weight loss, and cardiovascular uses. However, most of the MCFAs people use now are extracted from tropical plants, which is very time-consuming and limits the amount of medium chain triglyceride. Our goal is to strengthen the MCFAs synthesis pathway and optimize T7 initiation by over-expressing key enzymes in the reverse fatty acid β-oxidation cycle in E. coli to increase the production of MCFAs. We constructed pET28a-EGFP plasmid with fluorescent protein. In order to find the mutation that works the best, we designed point mutations C1, C2, C3, and C4 on T7promoter, and constructed these four T7promoter mutants on plasmids. We found out that the C1 mutation was the strongest promoter, which hooked up the selected mutations to the four enzymes that actually build the medium-chain fatty acids. This mutated promoter was then applied to the four enzymes used in our pathway to increase the amount of medium-chain fatty acids production.
