E. coli Engineered for Triglyceride Production.
Abstract: The goal of
this work was to engineer Escherichia coli to synthesize and accumulate triglycerides
(TG). Triglycerides, which are commonly sourced from plant oils, are substrates for
products such as biodiesel, lotions, lubricants and cooking oils. A liquid
chromatography-mass spectrometry (LCMS) method, developed to analyze the production of
triglycerides by the engineered E. coli s... read moretrains, demonstrated a yield of 1 mg/L
triglycerides with 2 g/L dry cell weight (DCW) in LB medium containing 5 g/L glucose.
Triglycerides were produced from cell wall phospholipid precursors through engineered
expression of two enzymes, PgpB and DGAT, which perform phosphatidic acid phosphatase
activity and diacylglycerol acyl transferase activity, respectively. In addition, E.
coli was engineered to express a third protein, phasin, which was hypothesized to
improve triglyceride accumulation through stabilizing effects on hydrophobic lipid
droplets in the cytoplasm.
Thesis (M.S.)--Tufts University, 2012.
Submitted to the Dept. of Chemical and Biological Engineering.
Advisors: Kyongbum Lee, and Blaine Pfeifer.
Committee: Kyongbum Lee, Blaine Pfeifer, and Qiaobing Xu.
Keywords: Cellular biology, Biochemistry, and Microbiology.read less