Engineering Escherichia coli for the production of adipic acid through the reversed β-oxidation pathway

Thirumalaisamy Babu; Eun Ju Yun; Sooah Kim; Do Hyoung Kim; Kwang Hyeon Liu; Soo Rin Kim; Kyoung Heon Kim

doi:10.1016/j.procbio.2015.09.018

Engineering Escherichia coli for the production of adipic acid through the reversed β-oxidation pathway

Thirumalaisamy Babu
, Eun Ju Yun
, Sooah Kim
, Do Hyoung Kim
, Kwang Hyeon Liu
, Soo Rin Kim^*
, Kyoung Heon Kim

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

36 Citations (Scopus)

Abstract

The current study aimed to construct a functionally reversed β-oxidation cycle in Escherichia coli for the production of adipic acid, one of the most important dicarboxylic acids. Starting from the condensation of acetyl-CoA and succinyl-CoA, a synthetic pathway mimicking a reversed β-oxidation cycle was constructed in E. coli to produce adipyl-CoA, consisting of four enzymatic steps. Products of the synthetic pathway were confirmed by liquid chromatography-tandem mass spectrometry. To convert adipyl-CoA into adipic acid, a putative adipyl-CoA thioesterase (encoded by Acot8 from Mus musculus) was selected, and its functional expression in E. coli was confirmed by gas chromatography-mass spectrometry. Finally, when the reversed β-oxidation cycle and Acot8 were co-expressed in E. coli, the resulting strain produced 12 μg/L of adipic acid from glucose. These results suggest that the synthetic adipic acid pathway functions in E. coli but requires further optimization for increased production of adipic acid.

Original language	English
Pages (from-to)	2066-2071
Number of pages	6
Journal	Process Biochemistry
Volume	50
Issue number	12
DOIs	https://doi.org/10.1016/j.procbio.2015.09.018
Publication status	Published - 2015 Dec 1

Bibliographical note

Funding Information:
This work was supported by grants from the National Research Foundation of Korea ( 2013M1A2A2072597 ) and the Advanced Biomass R&D Center of Korea ( 2011-0031353 ), both funded by the Korean Government (MSIP). This research was performed at the Korea University Food Safety Hall for the Institute of Biomedical Science and Food Safety.

Publisher Copyright:
© 2015 Elsevier Ltd. All rights reserved.

Copyright:
Copyright 2015 Elsevier B.V., All rights reserved.

Keywords

Acetyl-CoA
Acot8
Adipic acid
Dicarboxylic acid
Succinyl-CoA
β-Oxidation cycle

ASJC Scopus subject areas

Bioengineering
Biochemistry
Applied Microbiology and Biotechnology

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10.1016/j.procbio.2015.09.018

Cite this

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title = "Engineering Escherichia coli for the production of adipic acid through the reversed β-oxidation pathway",

abstract = "The current study aimed to construct a functionally reversed β-oxidation cycle in Escherichia coli for the production of adipic acid, one of the most important dicarboxylic acids. Starting from the condensation of acetyl-CoA and succinyl-CoA, a synthetic pathway mimicking a reversed β-oxidation cycle was constructed in E. coli to produce adipyl-CoA, consisting of four enzymatic steps. Products of the synthetic pathway were confirmed by liquid chromatography-tandem mass spectrometry. To convert adipyl-CoA into adipic acid, a putative adipyl-CoA thioesterase (encoded by Acot8 from Mus musculus) was selected, and its functional expression in E. coli was confirmed by gas chromatography-mass spectrometry. Finally, when the reversed β-oxidation cycle and Acot8 were co-expressed in E. coli, the resulting strain produced 12 μg/L of adipic acid from glucose. These results suggest that the synthetic adipic acid pathway functions in E. coli but requires further optimization for increased production of adipic acid.",

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note = "Funding Information: This work was supported by grants from the National Research Foundation of Korea ( 2013M1A2A2072597 ) and the Advanced Biomass R\&D Center of Korea ( 2011-0031353 ), both funded by the Korean Government (MSIP). This research was performed at the Korea University Food Safety Hall for the Institute of Biomedical Science and Food Safety. Publisher Copyright: {\textcopyright} 2015 Elsevier Ltd. All rights reserved. Copyright: Copyright 2015 Elsevier B.V., All rights reserved.",

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AU - Liu, Kwang Hyeon

AU - Kim, Soo Rin

AU - Kim, Kyoung Heon

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PY - 2015/12/1

Y1 - 2015/12/1

N2 - The current study aimed to construct a functionally reversed β-oxidation cycle in Escherichia coli for the production of adipic acid, one of the most important dicarboxylic acids. Starting from the condensation of acetyl-CoA and succinyl-CoA, a synthetic pathway mimicking a reversed β-oxidation cycle was constructed in E. coli to produce adipyl-CoA, consisting of four enzymatic steps. Products of the synthetic pathway were confirmed by liquid chromatography-tandem mass spectrometry. To convert adipyl-CoA into adipic acid, a putative adipyl-CoA thioesterase (encoded by Acot8 from Mus musculus) was selected, and its functional expression in E. coli was confirmed by gas chromatography-mass spectrometry. Finally, when the reversed β-oxidation cycle and Acot8 were co-expressed in E. coli, the resulting strain produced 12 μg/L of adipic acid from glucose. These results suggest that the synthetic adipic acid pathway functions in E. coli but requires further optimization for increased production of adipic acid.

AB - The current study aimed to construct a functionally reversed β-oxidation cycle in Escherichia coli for the production of adipic acid, one of the most important dicarboxylic acids. Starting from the condensation of acetyl-CoA and succinyl-CoA, a synthetic pathway mimicking a reversed β-oxidation cycle was constructed in E. coli to produce adipyl-CoA, consisting of four enzymatic steps. Products of the synthetic pathway were confirmed by liquid chromatography-tandem mass spectrometry. To convert adipyl-CoA into adipic acid, a putative adipyl-CoA thioesterase (encoded by Acot8 from Mus musculus) was selected, and its functional expression in E. coli was confirmed by gas chromatography-mass spectrometry. Finally, when the reversed β-oxidation cycle and Acot8 were co-expressed in E. coli, the resulting strain produced 12 μg/L of adipic acid from glucose. These results suggest that the synthetic adipic acid pathway functions in E. coli but requires further optimization for increased production of adipic acid.

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Engineering Escherichia coli for the production of adipic acid through the reversed β-oxidation pathway

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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