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Enhancing pentose phosphate pathway in Corynebacterium glutamicum to improve L-isoleucine production

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논문

Enhancing pentose phosphate pathway in Corynebacterium glutamicum to improve L-isoleucine production

학술지

Biotechnology and applied biochemistry

저자명

Ma, Wenjian; Wang, Jianli; Li, Ye; Hu, Xiaoqing; Shi, Feng; Wang, Xiaoyuan

초록

<P><B>Abstract</B></P><P>Three genes, <I>gnd</I>, <I>pgl</I>, and <I>fbp</I>, relevant to the pentose phosphate pathway (PPP) were overexpressed in <I>Corynebacterium glutamicum</I> IWJ001, leading to increase of l&#8208;isoleucine production. The transcriptional levels of <I>gnd</I>, <I>pgl</I>, and <I>fbp</I> significantly increased in IWJ001/pDXW&#8208;8&#8208;<I>gnd</I>&#8208;<I>fbp</I>&#8208;<I>pgl</I>. Compared with the control strain IWJ001/pDXW&#8208;8, intracellular NADPH/NADP<SUP>+</SUP> ratios in IWJ001/pDXW&#8208;8&#8208;<I>gnd</I> and IWJ001/pDXW&#8208;8&#8208;<I>gnd</I>&#8208;<I>fbp</I> cells grown for 36 H increased threefold and fourfold, respectively, indicating that overexpression of <I>gnd</I> and <I>fbp</I> redirected the carbon flux to PPP. Intracellular NADPH/NADP<SUP>+</SUP> ratio in IWJ001/pDXW&#8208;8&#8208;<I>gnd</I>&#8208;<I>fbp</I>&#8208;<I>pgl</I> grown for 36 H was similar to IWJ001/pDXW&#8208;8, suggesting that the NADPH produced by PPP could be quickly consumed for l&#8208;isoleucine production. 10.9 and 28.96 g/L of l&#8208;isoleucine was produced in IWJ001/pDXW&#8208;8&#8208;<I>gnd</I>&#8208;<I>fbp</I>&#8208;<I>pgl</I> in shake flask cultivation and fed&#8208;batch fermentation, respectively. In addition, IWJ001/pDXW&#8208;8&#8208;<I>gnd</I>&#8208;<I>fbp</I>&#8208;<I>pgl</I> grew fast, its dry cell weight reached 49 g/L after 48 H, whereas the start strain IWJ001/pDXW&#8208;8 reached only 40 g/L. After 96 H fermentation, l&#8208;isoleucine yield on glucose in IWJ001/pDXW&#8208;8&#8208;<I>gnd</I>&#8208;<I>fbp</I>&#8208;<I>pgl</I> reached 0.138 g/g. The results demonstrate that carbon flux redirection to PPP is an efficient approach to enhance l&#8208;isoleucine production in <I>C. glutamicum</I>.</P>

발행연도

2016

ISSN

0885-4513

63

6

페이지

pp.877-885

주제어

branched&#x2010; chain amino acids; C. glutamicum; l&#x2010; isoleucine; NADPH; pentose phosphate pathway;

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1 2023-12-11

논문; 2016-12-31

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