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Efficient production of xylitol by the integration of multiple copies of xylose reductase gene and the deletion of Embden–Meyerhof–Parnas pathway-associated genes to enhance NADPH regeneration in Escherichia coli

  • Metabolic Engineering and Synthetic Biology - Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Cofactor supply is a rate-limiting step in the bioconversion of xylose to xylitol. Strain WZ04 was first constructed by a novel simultaneous deletion–insertion strategy, replacing ptsG, xylAB and ptsF in wild-type Escherichia coli W3110 with three mutated xylose reductase genes (xr) from Neurospora crassa. Then, the pfkA, pfkB, pgi and/or sthA genes were deleted and replaced by xr to investigate the influence of carbon flux toward the pentose phosphate pathway and/or transhydrogenase activity on NADPH generation. The deletion of pfkA/pfkB significantly improved NADPH supply, but minimally influenced cell growth. The effects of insertion position and copy number of xr were examined by a quantitative real-time PCR and a shake-flask fermentation experiment. In a fed-batch fermentation experiment with a 15-L bioreactor, strain WZ51 produced 131.6 g L−1 xylitol from hemicellulosic hydrolysate (xylitol productivity: 2.09 g L−1 h−1). This study provided a potential approach for industrial-scale production of xylitol from hemicellulosic hydrolysate.

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Acknowledgements

This work was supported by National Natural Science Foundation of China (21376215), National Basic Research Program of China (2011CB710803) and National High Technology Research and Development Program of China (2012AA022302).

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  1. Xinsong Yuan and Jiping Wang contributed equally to this study.

Authors and Affiliations

  1. Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, 310027, People’s Republic of China

    Xinsong Yuan, Jiping Wang, Jianping Lin, Lirong Yang & Mianbin Wu

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  1. Xinsong Yuan
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  2. Jiping Wang
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  3. Jianping Lin
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Correspondence to Mianbin Wu.

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Yuan, X., Wang, J., Lin, J. et al. Efficient production of xylitol by the integration of multiple copies of xylose reductase gene and the deletion of Embden–Meyerhof–Parnas pathway-associated genes to enhance NADPH regeneration in Escherichia coli. J Ind Microbiol Biotechnol 46, 1061–1069 (2019). https://doi.org/10.1007/s10295-019-02169-3

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  • DOI: https://doi.org/10.1007/s10295-019-02169-3

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