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High-level intra- and extra-cellular production of d-psicose 3-epimerase via a modified xylose-inducible expression system in Bacillus subtilis

  • Genetics and Molecular Biology of Industrial Organisms - Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

d-Psicose 3-epimerase (DPEase) converts d-fructose into d-psicose which exists in nature in limited quantities and has key physiological functions. In this study, RDPE (DPEase from Ruminococcus sp. 5_1_39BFAA) was successfully constitutively expressed in Bacillus subtilis, which is the first report of its kind. Three sugar-inducible promoters were compared, and the xylose-inducible promoter P xylA was proved to be the most efficient for RDPE production. Based on the analysis of the inducer concentration and RDPE expression, we surmised that there was an extremely close correlation between the intracellular RDPE expression and xylose accumulation level. Subsequently, after the metabolic pathway of xylose was blocked by deletion of xylAB, the intra- and extra-cellular RDPE expression was significantly enhanced. Meanwhile, the optimal xylose induction concentration was reduced from 4.0 to 0.5 %. Eventually, the secretion level of RDPE reached 95 U/mL and 2.6 g/L in a 7.5-L fermentor with the fed-batch fermentation, which is the highest production of DPEase by a microbe to date.

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Acknowledgments

Authors would like to express their thanks to financial supports from National Nature Science Foundation of China (31370089, 21506244, 31570303), State Key Development 973 Program for Basic Research of China (2013CB733601), Natural Science Foundation of Liaoning Province of China (2014026012) and Tianjin Nature Science Foundation (16JCYBJC23500, 15JCQNJC09500).

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Authors and Affiliations

  1. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, People’s Republic of China

    Jingqi Chen, Yueming Zhu, Gang Fu, Yafeng Song, Yuanxia Sun & Dawei Zhang

  2. Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, People’s Republic of China

    Jingqi Chen, Yafeng Song & Dawei Zhang

  3. National Engineering Laboratory for Industrial Enzymes, Tianjin, 300308, People’s Republic of China

    Yueming Zhu, Gang Fu, Yuanxia Sun & Dawei Zhang

  4. School of Biological Engineering, Dalian Polytechnic University, Dalian, 116034, People’s Republic of China

    Zhaoxia Jin

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  1. Jingqi Chen
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Correspondence to Yuanxia Sun or Dawei Zhang.

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Chen, J., Zhu, Y., Fu, G. et al. High-level intra- and extra-cellular production of d-psicose 3-epimerase via a modified xylose-inducible expression system in Bacillus subtilis . J Ind Microbiol Biotechnol 43, 1577–1591 (2016). https://doi.org/10.1007/s10295-016-1819-6

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  • DOI: https://doi.org/10.1007/s10295-016-1819-6

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