Food and Fermentation Industries >

2024 , Vol. 50 >Issue 12: 17 - 23

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.036616

Effect of C/N terminus and “Thumb” structure of recombinant xylanase XynH on its catalytic properties

  • ZHU Aodi ,
  • ZHAO Ting ,
  • XU Jin ,
  • CHEN Maobin ,
  • FANG Shangling ,
  • LI Qin
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  • 1(Cooperative Innovation Center of Industrial Fermentation (Ministry of Education & Hubei Province), Key Laboratory of Fermentation Engineering (Ministry of Education), Hubei Key Laboratory of Industrial Microbiology, School of Food and Biological Engineering, Hubei University of Technology, Wuhan 430068, China)
    2(College of Food Science, Sichuan Agricultural University, Ya′an 625014, China)

Received date: 2023-06-29

  Revised date: 2023-08-07

  Online published: 2024-07-11

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Abstract

Xylanase is widely used in papermaking, feed, energy, food, medicine, and other industries.It is of great significance to use genetic engineering technology to quickly tap xylanase resources suitable for industrial production.Based on bioinformatics analysis, we successfully constructed mutant X-N, X-C, and X-f based on molecular modification of the C/N terminal and “Thumb” structure of xylanase XynH.Compared with the proenzyme XynH, the optimal pH of X-N was 5.5, and the other enzymes showed the highest enzyme activity at pH 5.0.The residual enzyme activity of all mutants and XynH at pH 4.0-6.0 was about 60%, and they had good acid resistance.The optimum temperature of X-N (60 ℃) was increased by 5 ℃ compared with other enzymes.After heat preservation at 55 ℃ for 1 h, the residual enzyme activity of X-N and X-f was higher than that of XynH, and the heat resistance of X-N was better.All the mutants showed weak affinity to beech xylan, but showed higher conversion rate.In this paper, the molecular modification of xylanase by genetic engineering means was used to construct three mutants, all of which showed good acid resistance, the optimal pH of X-N was slightly higher than that of the original enzyme, and the stability was better at high temperature.N-terminal replacement improves the thermal stability of xylanase, and C-terminal replacement may involve a part of the catalytic domain, resulting in a change in enzyme activity.The modification of the end of "Thumb" may narrow the cracks at the active site, preventing the entry of the substrate, resulting in a reduction in the catalytic efficiency of the enzyme.The difference between the mutant and the original enzyme in enzymology and the relationship between structure and function revealed by the mutant provide some ideas and strategies for the improvement and application of xylanase.

Key words： xylanase; bioinformatics; molecular modification; enzymatic property

Cite this article

ZHU Aodi , ZHAO Ting , XU Jin , CHEN Maobin , FANG Shangling , LI Qin . Effect of C/N terminus and “Thumb” structure of recombinant xylanase XynH on its catalytic properties[J]. Food and Fermentation Industries, 2024 , 50(12) : 17 -23 . DOI: 10.13995/j.cnki.11-1802/ts.036616

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