High-temperature-resistant beta-mannase mutant and preparation method thereof

By substituting specific amino acids for β-mannanase, a high temperature resistant mutant was obtained, which solved the problem of poor heat resistance of existing enzymes, achieved high enzyme activity under longer heating time under alkaline conditions, and expanded its application range.

CN119662604AActive Publication Date: 2025-03-21NANJING UNIV
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Patent Information

Application Number
CN202411925108.1
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2024-12-25
Publication Date
2025-03-21
Estimated Expiration
2044-12-25

AI Technical Summary

Technical Problem

The existing β-mannanase has poor heat resistance during production, transportation and application, resulting in unstable biological activity, which limits its application scope.

Method used

By performing specific amino acid replacement on the amino acid sequence of β-mannanase of the β-mannanase of the gray-capita β-mannanase, a high-temperature resistant β-mannanase mutant was obtained, and the specific replacement includes N381K, R123Y, M439L, etc., improving the thermal stability of the enzyme.

Benefits of technology

Under alkaline conditions, this mutant can maintain good enzyme activity under longer heating time. Compared with the original enzyme, high temperature treatment of 90°C in a buffer with pH 10.0 for 2 hours, the enzyme activity was increased by 89%, and it has broad application prospects.

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Abstract

The invention discloses a high temperature resistant beta-mannase mutant and a preparation method thereof, wild type beta-mannase comes from Coprinus cinerea, the wild type beta-mannase is modified by using a directed evolution means to obtain the high temperature resistant beta-mannase mutant, and the mutation of the mutant comprises at least one of N381K, R123Y and M439L. The beta-mannase mutant has good thermal stability under the alkaline condition and longer heating time, the enzyme activity of the mutant is improved by 107.0% compared with that of an original enzyme when the beta-mannase mutant is treated for 2 h at the high temperature of 90 DEG C in a buffer solution with the pH value of 10, the capacity of catalyzing beta-mannase hydrolysis by the enzyme mutant is obviously improved, and the beta-mannase mutant has wide application prospects.
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