Food and Fermentation Industries >

2022 , Vol. 48 >Issue 13: 16 - 24

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.029933

Optimization of fermentation conditions of recombinant Escherichia coli expressing haloalkane dehalogenase

  • MING Xiaodong ,
  • GUO Zitao ,
  • CHEN Jianxiong ,
  • GU Zhenghua ,
  • XIN Yu ,
  • SUN Haiyan ,
  • GUAN Yanming ,
  • ZHANG Liang
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  • 1(National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China)
    2(Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, China)
    3(China National Research Institute of Food and Fermentation Industries, Beijing, 100015, China)

Received date: 2021-11-02

  Revised date: 2021-11-28

  Online published: 2022-08-03

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Abstract

In this study, the fermentation of a recombinant Escherichia coli BL21(DE3)/pET28a-DhaA was optimized to improve its production of haloalkane dehalogenase. Firstly, based on TB medium, the components of culture medium were optimized at shaking flask level by single factor experiment and response surface experiment. Secondly, the fermentation conditions were optimized base on the optimal culture medium. Finally, the preliminary amplification was verified in a 5 L fermentation tank. The results showed that the optimal medium composition was glycerol 10 g/L, yeast powder 23 g/L, peptone 14 g/L, MgSO4·7H2O 1.3 g/L, ZnSO4·7H2O 0.1 g/L. The enzyme activity can reach (1 182.94±10.86) U/L, which was 94% higher than that of the initial medium. The optimal fermentation conditions were: the inoculum volume 5%, the loading volume 40 mL/250 mL, 0.4 mmol/L IPTG was added to induce 22 h at 20 ℃, when the OD600 reached 1.8 at 37 ℃. The enzyme activity was achieved at (3 585.83±15.02) U/L, which was 5.87 times higher than the initial conditions. At last, the enzyme activity reached (9 682.62±191.16) U/L in a 5 L fermentation tank,which was 2.7 times higher than that of a shaking flask. After optimization, the enzyme activity of haloalkane dehalogenase was greatly improved, and our study provides a foundation for scale-up production.

Key words： haloalkane dehalogenase; Escherichia coli; fermentation optimization; response surface method

Cite this article

MING Xiaodong , GUO Zitao , CHEN Jianxiong , GU Zhenghua , XIN Yu , SUN Haiyan , GUAN Yanming , ZHANG Liang . Optimization of fermentation conditions of recombinant Escherichia coli expressing haloalkane dehalogenase[J]. Food and Fermentation Industries, 2022 , 48(13) : 16 -24 . DOI: 10.13995/j.cnki.11-1802/ts.029933

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