Amide hydrolase mutant and application thereof

The L447A mutation of the amide hydrolase RsAD solved the problem of low amide hydrolase activity, significantly improving the yield and catalytic efficiency of indole acetic acid, especially showing higher activity during large-volume substrate reactions.

CN119709703AActive Publication Date: 2025-03-28INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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Patent Information

Application Number
CN202411642866.2
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2024-11-18
Publication Date
2025-03-28
Estimated Expiration
2044-11-18

AI Technical Summary

Technical Problem

The existing amide hydrolase activity is low, resulting in the hydrolysis of indole acetamide to produce indole acetic acid, especially in the low catalytic efficiency when reacting with large volume substrates.

Method used

By mutating the amide hydrolase RsAD derived from the microorganism Rhodococcus sp., especially mutating leucine at position 447 to alanine, the amide hydrolase mutant L447A was constructed to enhance its enzyme activity to increase the yield of indole acetic acid.

Benefits of technology

The enzyme activity of the mutant L447A was significantly improved, the yield of indole acetic acid was greatly improved, and the catalytic efficiency was significantly improved, especially when it showed higher activity when reacting with large volume substrates.

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Abstract

The invention relates to the technical field of enzyme engineering, synthetic biology and microbial fermentation, in particular to an amide hydrolase mutant and application thereof. The substrate range of the amide hydrolase RsAD from the microorganism Rhodococcus sp. Is relatively narrow, and after the amino acid residue L447 in the substrate channel of the amide hydrolase RsAD is mutated into alanine, the reaction enzyme activity of the alanine on the large-volume substrate, namely indoleacetamide, is increased, so that the production efficiency of indoleacetic acid is greatly improved.
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