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.
| Original language | English |
|---|---|
| Pages (from-to) | 44-51 |
| Number of pages | 8 |
| Journal | Enzyme and Microbial Technology |
| Volume | 49 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 2011 Jun 10 |
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Keywords
- Acetyl-CoA
- E. coli MG1655
- Fatty acid biosynthesis
- Fatty acyl-ACP chain
- Long chain fatty acid
- Malonyl-[acp]
- Malonyl-CoA
ASJC Scopus subject areas
- Biochemistry
- Biotechnology
- Applied Microbiology and Biotechnology
Cite this
Development of Escherichia coli MG1655 strains to produce long chain fatty acids by engineering fatty acid synthesis (FAS) metabolism. / Jeon, Eunyoung; Lee, Sunhee; Won, Jong In; Han, Sung Ok; Kim, Jihyeon; Lee, Jinwon.
In: Enzyme and Microbial Technology, Vol. 49, No. 1, 10.06.2011, p. 44-51.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Development of Escherichia coli MG1655 strains to produce long chain fatty acids by engineering fatty acid synthesis (FAS) metabolism
AU - Jeon, Eunyoung
AU - Lee, Sunhee
AU - Won, Jong In
AU - Han, Sung Ok
AU - Kim, Jihyeon
AU - Lee, Jinwon
PY - 2011/6/10
Y1 - 2011/6/10
N2 - 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.
AB - 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.
KW - Acetyl-CoA
KW - E. coli MG1655
KW - Fatty acid biosynthesis
KW - Fatty acyl-ACP chain
KW - Long chain fatty acid
KW - Malonyl-[acp]
KW - Malonyl-CoA
UR - http://www.scopus.com/inward/record.url?scp=79957546202&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=79957546202&partnerID=8YFLogxK
U2 - 10.1016/j.enzmictec.2011.04.001
DO - 10.1016/j.enzmictec.2011.04.001
M3 - Article
VL - 49
SP - 44
EP - 51
JO - Enzyme and Microbial Technology
JF - Enzyme and Microbial Technology
SN - 0141-0229
IS - 1
ER -