Collagenase mutant, gene segment, recombinant plasmid, recombinant expression system and application thereof, and method for preparing collagenase mutant and collagen tripeptide

By efficiently secreting and expressing collagenase mutant KT231 in Streptomyces S. lividansTK24 and cooperating with papaya chymoprotein, the problems of low enzyme activity and high production cost in the existing collagenase production methods are solved, and collagen tripeptide is efficiently prepared, suitable for a variety of skin and joint health products.

CN119776327AActive Publication Date: 2025-04-08HUBEI KAITAI BIOTECHNOLOGY CO LTD
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Patent Information

Application Number
CN202510292428.6
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-03-12
Publication Date
2025-04-08
Estimated Expiration
2045-03-12

AI Technical Summary

Technical Problem

The existing collagenase production methods have complex processes, high costs and potential disease transmission risks, and microbial fermentation methods face the problems of low enzyme activity and high production costs in practical applications.

Method used

A collagenase mutant KT231 that achieves efficient secretion and expression in Streptomyces S. lividansTK24 was developed, and the proportion of collagen tripeptides in the fish skin enzymatic solution was increased through complexing with commercially available papaya chymoproteinase.

Benefits of technology

Collagenase mutant KT231 is highly effective in enzymatic decomposition of cod fish skin at 60°C. It has excellent heat resistance and high enzyme activity, which significantly increases the proportion of collagen tripeptide in the enzymatic decomposition of fish skin, and has no bitter taste. It is suitable for the preparation of anti-wrinkle moisturizing, repairing damaged skin and other products.

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Abstract

The invention discloses a collagenase mutant, a gene segment, a recombinant plasmid, a recombinant expression system and application thereof, and a method for preparing the collagenase mutant and collagen tripeptide, and belongs to the field of gene engineering and fermentation engineering. The invention provides a collagenase mutant KT231 capable of realizing high-efficiency secretory expression in streptomyces S.lividans TK24 and a preparation method of the collagenase mutant KT231. The collagenase mutant KT231 can be used for efficiently carrying out enzymolysis on the skin of the narrow gadus at 60 DEG C, has excellent heat resistance and higher enzyme activity, and can be used for efficiently preparing the collagen tripeptide from the skin of the narrow gadus. The collagen peptide is compounded with commercially available papaya chymotrypsin, so that the proportion of collagen tripeptide in the fish skin enzymatic hydrolysate can be remarkably increased, and the fish skin enzymatic hydrolysate has no bitter taste. The method has wide application prospects in preparation of products, tissue engineering materials and wound healing accelerators for resisting wrinkles, preserving moisture, repairing damaged skin and improving bone and joint health.
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