Development of an orthogonal fatty acid biosynthesis system in E. coli for oleochemical production

Metab Eng. 2015 Jul;30:1-6. doi: 10.1016/j.ymben.2015.04.003. Epub 2015 Apr 15.

Abstract

Here we report recombinant expression and activity of several type I fatty acid synthases that can function in parallel with the native Escherichia coli fatty acid synthase. Corynebacterium glutamicum FAS1A was the most active in E. coli and this fatty acid synthase was leveraged to produce oleochemicals including fatty alcohols and methyl ketones. Coexpression of FAS1A with the ACP/CoA-reductase Maqu2220 from Marinobacter aquaeolei shifted the chain length distribution of fatty alcohols produced. Coexpression of FAS1A with FadM, FadB, and an acyl-CoA-oxidase from Micrococcus luteus resulted in the production of methyl ketones, although at a lower level than cells using the native FAS. This work, to our knowledge, is the first example of in vivo function of a heterologous fatty acid synthase in E. coli. Using FAS1 enzymes for oleochemical production have several potential advantages, and further optimization of this system could lead to strains with more efficient conversion to desired products. Finally, functional expression of these large enzyme complexes in E. coli will enable their study without culturing the native organisms.

Keywords: Fatty acid synthase; Fatty alcohols; Methyl ketones; Synthetic biology.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacterial Proteins / biosynthesis*
  • Bacterial Proteins / genetics
  • Corynebacterium glutamicum / enzymology
  • Corynebacterium glutamicum / genetics*
  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Fatty Acid Synthases / biosynthesis*
  • Fatty Acid Synthases / genetics
  • Fatty Acids / biosynthesis*
  • Fatty Acids / genetics
  • Marinobacter / enzymology
  • Marinobacter / genetics*
  • Micrococcus luteus / enzymology
  • Micrococcus luteus / genetics*

Substances

  • Bacterial Proteins
  • Fatty Acids
  • Fatty Acid Synthases