The goal of this research was to develop recombinant Escherichia coli to improve fatty acid synthesis (FAS). Genes encoding acetyl-CoA carboxylase (accA, accB, accC), malonyl-CoA-[acyl-carrier-protein] transacylase (fabD), and acyl-acyl carrier protein thioesterase (EC 188.8.131.52 gene), which are all enzymes that catalyze key steps in the synthesis of fatty acids, were cloned and over-expressed in E. coli MG1655. The acetyl-CoA carboxylase (ACC) enzyme catalyzes the addition of CO 2 to acetyl-CoA to generate malonyl-CoA. The enzyme encoded by the fabD gene converts malonyl-CoA to malonyl-[acp], and the EC 184.108.40.206 gene converts fatty acyl-ACP chains to long chain fatty acids. All the genes except for the EC 220.127.116.11 gene were homologous to E. coli genes and were used to improve the enzymatic activities to over-express components of the FAS pathway through metabolic engineering. All recombinant E. coli MG1655 strains containing various gene combinations were developed using the pTrc99A expression vector. To observe changes in metabolism, the in vitro metabolites and fatty acids produced by the recombinants were analyzed. The fatty acids (C16) from recombinant strains were produced 1.23-2.41 times higher than that from the wild type.
Bibliographical noteFunding Information:
This work was supported by the New and Renewable Energy development of technology project funded by the Korean Government (Ministry of Knowledge Economy) (No. 2008-N-BI08-P-03 ).
- E. coli MG1655
- Fatty acid biosynthesis
- Fatty acyl-ACP chain
- Long chain fatty acid
ASJC Scopus subject areas
- Applied Microbiology and Biotechnology