Food and Fermentation Industries >

2021 , Vol. 47 >Issue 19: 50 - 56

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.026926

Improving the thermostability of Bacillus subtilis keratinase by rational design

  • MIAO Zhoudi ,
  • CHEN Xiwen ,
  • PENG Zheng ,
  • MAO Xinzhe ,
  • DU Guocheng ,
  • ZHANG Juan
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  • 1(Key Laboratory of Industrial Biotechnology,Ministry of Education,Jiangnan University,Wuxi 214122,China)
    2(School of Biotechnology,Jiangnan University,Wuxi 214122,China)
    3(Synergetic Innovation Center,Jiangnan University,Wuxi 214122,China)
    4(Key Laboratory of Carbohydrate Chemistry and Biotechnology,Ministry of Education,Jiangnan University,Wuxi 214122,China)

Received date: 2021-01-29

  Revised date: 2021-03-19

  Online published: 2021-11-04

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Abstract

Bacillus subtilis keratinase (KerZ1) could effectively degrade keratin waste at 60 ℃, however, its thermostability is not preferable. The rational design method was applied to increase its thermostability. First, the B-factor analysis and molecular dynamics simulation software AMBER16 were used to predict the flexible loop regions. Subsequently, combined with the β-turn residue statistic, the residues at the introduced position were replaced with conserved amino acids. The results showed that the half-life (t1/2) and half inactivation temperature (T50) of the A128D mutant increased by 12.32 min and 5.8 ℃, respectively. And the catalytic efficiency was not significantly affected. In addition, the analysis of the distribution of mutant residues and surrounding amino acid residues indicated that the increased surface hydrophilicity of the A128D mutant might be the main reason for thermostability. In conclusion, this strategy improves the thermostability of keratinase by modifying the flexible loop region, and lays the foundation for the industrial application of keratinase.

Key words： keratinase; thermostability; rational design method; B-factor; molecular dynamics simulation; β-turn

Cite this article

MIAO Zhoudi , CHEN Xiwen , PENG Zheng , MAO Xinzhe , DU Guocheng , ZHANG Juan . Improving the thermostability of Bacillus subtilis keratinase by rational design[J]. Food and Fermentation Industries, 2021 , 47(19) : 50 -56 . DOI: 10.13995/j.cnki.11-1802/ts.026926

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