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Metab Eng. 2019 Jan;51:99-109. doi: 10.1016/j.ymben.2018.08.007. Epub 2018 Aug 23.

Metabolic engineering of Corynebacterium glutamicum for the production of glutaric acid, a C5 dicarboxylic acid platform chemical.

Author information

1
Bio-based Chemistry Research Center, Advanced Convergent Chemistry Division, Korea Research Institute of Chemical Technology, P.O. Box 107, 141 Gajeong-ro, Yuseong-gu, Daejeon 34114 Republic of Korea.
2
Bio-based Chemistry Research Center, Advanced Convergent Chemistry Division, Korea Research Institute of Chemical Technology, P.O. Box 107, 141 Gajeong-ro, Yuseong-gu, Daejeon 34114 Republic of Korea; Department of Chemical and Biological Engineering, Korea University, 145 Anam-ro, Seongbuk-gu, Seoul 02841, Republic of Korea.
3
Division of Chemical Engineering and Materials Science, Ewha Womans University, 52 Ewhayeodae-gil, Seodaemun-gu, Seoul 03760, Republic of Korea.
4
Bio-based Chemistry Research Center, Advanced Convergent Chemistry Division, Korea Research Institute of Chemical Technology, P.O. Box 107, 141 Gajeong-ro, Yuseong-gu, Daejeon 34114 Republic of Korea; Department of Biological and Chemical Engineering, Hongik University, 2639 Sejong-ro, Sinan-ri, Jochiwon-eup, Sejong-si 30016, Republic of Korea.
5
Department of Biological and Chemical Engineering, Hongik University, 2639 Sejong-ro, Sinan-ri, Jochiwon-eup, Sejong-si 30016, Republic of Korea.
6
Department of Chemical and Biological Engineering, Korea University, 145 Anam-ro, Seongbuk-gu, Seoul 02841, Republic of Korea.
7
Metabolic Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 Plus program), KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea.
8
Department of Chemical and Biomolecular Engineering (BK21 Plus program), KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea.
9
Division of Chemical Engineering and Materials Science, Ewha Womans University, 52 Ewhayeodae-gil, Seodaemun-gu, Seoul 03760, Republic of Korea. Electronic address: parksj93@ewha.ac.kr.
10
Bio-based Chemistry Research Center, Advanced Convergent Chemistry Division, Korea Research Institute of Chemical Technology, P.O. Box 107, 141 Gajeong-ro, Yuseong-gu, Daejeon 34114 Republic of Korea. Electronic address: jcjoo@krict.re.kr.

Abstract

Corynebacterium glutamicum was metabolically engineered for the production of glutaric acid, a C5 dicarboxylic acid that can be used as platform building block chemical for nylons and plasticizers. C. glutamicum gabT and gabD genes and Pseudomonas putida davT and davD genes encoding 5-aminovalerate transaminase and glutarate semialdehyde dehydrogenase, respectively, were examined in C. glutamicum for the construction of a glutaric acid biosynthesis pathway along with P. putida davB and davA genes encoding lysine 2-monooxygenase and delta-aminovaleramidase, respectively. The glutaric acid biosynthesis pathway constructed in recombinant C. glutamicum was engineered by examining strong synthetic promoters PH30 and PH36, C. glutamicum codon-optimized davTDBA genes, and modification of davB gene with an N-terminal His6-tag to improve the production of glutaric acid. It was found that use of N-terminal His6-tagged DavB was most suitable for the production of glutaric acid from glucose. Fed-batch fermentation using the final engineered C. glutamicum H30_GAHis strain, expressing davTDA genes along with davB fused with His6-tag at N-terminus could produce 24.5 g/L of glutaric acid with low accumulation of l-lysine (1.7 g/L), wherein 5-AVA accumulation was not observed during fermentation.

KEYWORDS:

Codon optimization; Corynebacterium glutamicum; Fed-batch fermentation; Glutaric acid; His(6)-tag; L‐Lysine; davTDBA

PMID:
30144560
DOI:
10.1016/j.ymben.2018.08.007
[Indexed for MEDLINE]

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