Food and Fermentation Industries >

2023 , Vol. 49 >Issue 2: 47 - 53

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.031405

Recombined expression and optimized modification of alginate lyase from Isoptericola halotolerans

  • WU Wen ,
  • ZHU Fengshuai ,
  • YU Jiale ,
  • LI Heng ,
  • GONG Jinsong ,
  • XU Zhenghong ,
  • SHI Jinsong
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  • 1(Key Laboratory of the Ministry of Education of Sugar Chemistry and Biotechnology, College of Life Sciences and Health Engineering, Jiangnan University, Wuxi 214122, China)
    2(National Engineering Research Center for Food Fermentation and Food Biomanufacturing, Jiangnan University, Wuxi 214122, China)
    3(College of Bioengineering, Jiangnan University, Wuxi 214122, China)

Received date: 2022-03-04

  Revised date: 2022-04-05

  Online published: 2023-02-15

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Abstract

For further expanding the high-efficiency expression of alginate lyase in Bacillus subtilis, a segment of the alginate lyase gene named aly-ih from Isoptericola halotolerans WX was employed to construct an engineered strain of B. subtilis-pMA5-aly-ih. The catalytic activity of the enzyme was successively improved by the strategies of promoter modification and medium optimization. Herein, the enzymatic activity was increased by 4-fold compared with the initial enzyme through replacing the endogenous promoter HpaⅡ by mpr with homologous recombination method. The activity of the recombinase was further increased by 5.1-fold when the copy number of promoter mpr reached 3. The optimized fermentation medium was as following: 16 g/L glycerol, 16 g/L soy peptone, 25 mmol/L Mn2+. With the above conditions, the enzymatic activity of Aly-IH reached 320 U/mL which was 10.7 times as much as the initial enzyme. Aly-IH could efficiently degrade alginate to oligosaccharides with an average degree of polymerization of 2.3. Conclusively, the modified engineered Aly-IH in B. subtilis exerted prominent catalytic performance and potential application.

Key words： alginate lyase; Bacillus subtilis; promoter modification; media optimization

Cite this article

WU Wen , ZHU Fengshuai , YU Jiale , LI Heng , GONG Jinsong , XU Zhenghong , SHI Jinsong . Recombined expression and optimized modification of alginate lyase from Isoptericola halotolerans [J]. Food and Fermentation Industries, 2023 , 49(2) : 47 -53 . DOI: 10.13995/j.cnki.11-1802/ts.031405

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