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Construction of allitol synthesis pathway by multi-enzyme coexpression in Escherichia coli and its application in allitol production

  • Biocatalysis
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

An engineered strain for the conversion of d-fructose to allitol was developed by constructing a multi-enzyme coupling pathway and cofactor recycling system in Escherichia coli. d-Psicose-3-epimerase from Ruminococcus sp. and ribitol dehydrogenase from Klebsiella oxytoca were coexpressed to form the multi-enzyme coupling pathway for allitol production. The cofactor recycling system was constructed using the formate dehydrogenase gene from Candida methylica for continuous NADH supply. The recombinant strain produced 10.62 g/l allitol from 100 mM d-fructose. To increase the intracellular concentration of the substrate, the glucose/fructose facilitator gene from Zymomonas mobilis was incorporated into the engineered strain. The results showed that the allitol yield was enhanced significantly to 16.53 g/l with a conversion rate of 92 %. Through optimizing conversion conditions, allitol was produced effectively on a large scale by the whole-cell biotransformation system; the yield reached 48.62 g/l when 500 mM d-fructose was used as the substrate.

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Acknowledgments

This work was financially supported by the National Natural Science Foundation of China (31101303), the High Technology and Research Development Program (2013AA102102), and the Key Programs of the Chinese Academy of Sciences (KSZD-EW-Z-019).

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    Authors and Affiliations

    1. National Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Xiqi Road 32#, Tianjin Airport Economic Area, Tianjin, 300308, People’s Republic of China

      Yueming Zhu, Pingping Liu, Jiangang Yang, Xueli Zhang & Yuanxia Sun

    2. Key Laboratory of Cellular Physiology, Ministry of Education, Shanxi Medical University, Taiyuan, 030001, Shanxi, China

      Hongyi Li

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    1. Yueming Zhu
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    2. Hongyi Li
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    3. Pingping Liu
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    5. Xueli Zhang
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    Correspondence to Xueli Zhang.

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    Y. Zhu and H. Li have contributed equally to the work.

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    Zhu, Y., Li, H., Liu, P. et al. Construction of allitol synthesis pathway by multi-enzyme coexpression in Escherichia coli and its application in allitol production. J Ind Microbiol Biotechnol 42, 661–669 (2015). https://doi.org/10.1007/s10295-014-1578-1

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