期刊
BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY
卷 63, 期 6, 页码 877-885出版社
WILEY-BLACKWELL
DOI: 10.1002/bab.1442
关键词
branched-chain amino acids; C. glutamicum; L-isoleucine; NADPH; pentose phosphate pathway
资金
- National Natural Science Foundation of China [NSFC31370131]
- Six Talent Peaks Project of Jiangsu Province [2012-SWYY-008]
- Key Research Project of Jiangnan University [JUSRP51303A]
Three genes, gnd, pgl, and fbp, relevant to the pentose phosphate pathway (PPP) were overexpressed in Corynebacterium glutamicum IWJ001, leading to increase of L-isoleucine production. The transcriptional levels of gnd, pgl, and fbp significantly increased in IWJ001/pDXW-8-gnd-fbp-pgl. Compared with the control strain IWJ001/pDXW-8, intracellular NADPH/NADP(+) ratios in IWJ001/pDXW-8-gnd and IWJ001/pDXW-8-gnd-fbp cells grown for 36 H increased threefold and fourfold, respectively, indicating that overexpression of gnd and fbp redirected the carbon flux to PPP. Intracellular NADPH/NADP(+) ratio in IWJ001/pDXW-8-gnd-fbp-pgl grown for 36 H was similar to IWJ001/pDXW-8, suggesting that the NADPH produced by PPP could be quickly consumed for L-isoleucine production. 10.9 and 28.96 g/L of L-isoleucine was produced in IWJ001/pDXW-8-gnd-fbp-pgl in shake flask cultivation and fed-batch fermentation, respectively. In addition, IWJ001/pDXW-8-gnd-fbp-pgl grew fast, its dry cell weight reached 49 g/L after 48 H, whereas the start strain IWJ001/pDXW-8 reached only 40 g/L. After 96 H fermentation, L-isoleucine yield on glucose in IWJ001/pDXW-8-gnd-fbp-pgl reached 0.138 g/g. The results demonstrate that carbon flux redirection to PPP is an efficient approach to enhance L-isoleucine production in C. glutamicum. (C) 2015 International Union of Biochemistry and Molecular Biology, Inc. Volume 63, Number 6, Pages 877-885, 2016
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