期刊
METABOLIC ENGINEERING
卷 51, 期 -, 页码 99-109出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymben.2018.08.007
关键词
Glutaric acid; L-Lysine; Corynebacterium glutamicum; davTDBA; Codon optimization; His(6)-tag; Fed-batch fermentation
资金
- Mid-career Researcher Program through the National Research Foundation (NRF) of Korea - the Ministry of Science and ICT (MSIT) [NRF-2016R1A2B4008707]
- Technology Development Program to Solve Climate Changes on Systems Metabolic Engineering for Biorefineries from MSIT through the NRF of Korea [NRF-2015M1A2A2035810]
- Bio & Medical Technology Development Program MSIT through the NRF of Korea [NRF-2018M3A9H3020459]
- Lignin Biorefinery from MSIT through the NRF of Korea [NRF-2017M1A2A2087634]
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 His(6)-tag to improve the production of glutaric acid. It was found that use of N-terminal His(6)-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 His(6)-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.
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