Food and Fermentation Industries >

2022 , Vol. 48 >Issue 2: 224 - 230

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.027580

Improve the production efficiency of Bacillus natto nattokinase by knocking out the γ-PGA synthetic gene

  • HAN Yuxing ,
  • MENG Fanqiang ,
  • ZHOU Libang ,
  • LU Zhaoxin
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  • (College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095,China)

Received date: 2021-04-07

  Revised date: 2021-05-17

  Online published: 2022-02-28

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Abstract

Nattokinase is a thrombolytic enzyme produced by Bacillus natto. The polyglutamic acid (γ-PGA) produced during the cultivation of B. natto increases the viscosity of the fermentation broth, which makes the separation and purification of nattokinase difficult. It increases the cost and limits the large-scale production of nattokinase. In this study, the γ-PGA synthetic gene pgsB was knocked out by homologous recombination, to reduce the content of γ-PGA and the viscosity of the fermentation broth, which might improve the efficiency of separation and purification of nattokinase. The result showed that the yield of γ-PGA of pgsB-deleted strain reduced significantly compared to the wild strain. At the 24th hour of fermentation, the yield of γ-PGA of pgsB-deleted strain reduced 57.9%. Besides, there was no significant difference in cell density, indicating that the deletion of pgsB gene reduced the viscosity of the fermentation broth and had no effect on the growth of the bacteria. The recovery rate of nattokinase of the pgsB-deleted strain was increased by 19.2% in the process of purifying nattokinase by ultrafiltration. Therefore, the separation and purification efficiency of nattokinase had been improved after knocking out the pgsB gene, which might provide a new method for the industrial production of nattokinase.

Key words： double knockout; nattokinase; pgsB gene; γ-PGA

Cite this article

HAN Yuxing , MENG Fanqiang , ZHOU Libang , LU Zhaoxin . Improve the production efficiency of Bacillus natto nattokinase by knocking out the γ-PGA synthetic gene[J]. Food and Fermentation Industries, 2022 , 48(2) : 224 -230 . DOI: 10.13995/j.cnki.11-1802/ts.027580

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