title: Development of Escherichia coli MG1655 strains to produce long chain fatty acids by engineering fatty acid synthesis (FAS) metabolism.
abstract: 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 3.1.2.14 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 3.1.2.14 gene converts fatty acyl-ACP chains to long chain fatty acids. All the genes except for the EC 3.1.2.14 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.
authors: Department of Chemical and Biomolecular Engineering, Sogang University, Seoul 121-742, Republic of Korea.
authors: Eunyoung
authors: E
authors: Jeon
authors: Sunhee
authors: S
authors: Lee
authors: Jong-In
authors: JI
authors: Won
authors: Sung Ok
authors: SO
authors: Han
authors: Jihyeon
authors: J
authors: Kim
authors: Jinwon
authors: J
authors: Lee
mesh_terms: D000103:Acetyl-CoA Carboxylase / Q000235:genetics / Q000378:metabolism; D051080:Acyl-Carrier Protein S-Malonyltransferase / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D056804:Biofuels; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005227:Fatty Acids / Q000096:biosynthesis* / Q000737:chemistry; D005798:Genes, Bacterial; D008055:Lipids / Q000096:biosynthesis; D008314:Malonates / Q000378:metabolism; D060847:Metabolic Engineering / Q000379:methods*; D053858:Metabolic Networks and Pathways; D016296:Mutagenesis; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013869:Thiolester Hydrolases / Q000235:genetics / Q000378:metabolism
chemical_list: D056804:Biofuels; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D005227:Fatty Acids; D008055:Lipids; D008314:Malonates; D011994:Recombinant Proteins; C030290:malonic acid; D051080:Acyl-Carrier Protein S-Malonyltransferase; D013869:Thiolester Hydrolases; C018748:oleoyl-(acyl-carrier-protein) hydrolase; D000103:Acetyl-CoA Carboxylase