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Production of α‐ketobutyrate using engineered Escherichia coli via temperature shift
Author(s) -
Zhang Chenglin,
Qi Junsheng,
Li Yanjun,
Fan Xiaoguang,
Xu Qingyang,
Chen Ning,
Xie Xixian
Publication year - 2016
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.25959
Subject(s) - inducer , fermentation , threonine , biochemistry , escherichia coli , strain (injury) , bacteria , industrial fermentation , catabolism , dehydratase , generally recognized as safe , biology , enzyme , chemistry , food science , serine , gene , genetics , anatomy
Alpha‐ketobutyrate has been widely used in medicine and food additive industry. Because chemical and enzymatic methods are associated with many deficiencies, the recent focus shifted to fermentation for the production of α‐ketobutyrate. In this study, a genetically engineered strain THRDΔ rhtAΔilvIH/ pWSK29‐ ilvA was constructed, starting from an L‐threonine‐producing strain, by overexpressing threonine dehydratase (TD), reducing α‐ketobutyrate catabolism and L‐threonine export. The shake flask cultivation of THRDΔ rhtAΔilvIH/ pWSK29‐ ilvA allowed the production of 16.2 g/L α‐ketobutyrate. Accumulation of α‐ketobutyrate severely inhibited the cell growth. To develop a better TD expression system and avoid the usage of the expensive inducer IPTG, a temperature‐induced plasmid pBV220‐ ilvA was selected to transform the strain THRDΔ rhtAΔilvIH for α‐ketobutyrate production. The initial temperature was maintained at 35°C to guarantee normal cell growth, and then elevated to 40°C to induce the expression of TD. Under optimized conditions, the α‐ketobutyrate titer reached 40.8 g/L after 28 h of fermentation, with a productivity of 1.46 g/L/h and a yield of 0.19 g/g glucose, suggesting large‐scale production potential. Biotechnol. Bioeng. 2016;113: 2054–2059. © 2016 Wiley Periodicals, Inc.