Candida antarctica lipase B mutant and application thereof

By mutating Candida antarcticis lipase B to P143L/A284E and expressing it efficiently in Pichia pastoris, the problems of low catalytic activity and poor stability of Candida antarcticis lipase B were solved, achieving efficient synthesis of high-value-added products and reducing production costs.

CN122012457AActive Publication Date: 2026-05-12HANG ZHOU HE TAN CHUANG WU KE JI YOU XIAN GONG SI +3
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Patent Information

Application Number
CN202610495365.9
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2026-04-15
Publication Date
2026-05-12
Estimated Expiration
2046-04-15

AI Technical Summary

Technical Problem

The existing Candida antarctic lipase B has low catalytic activity, which makes it difficult to meet the synthesis requirements of high value-added products. In addition, it has poor stability in non-aqueous reaction systems and low expression levels, resulting in high costs for downstream separation and purification.

Method used

A mutant of Candida antarcticus lipase B (EXP01) was constructed by mutating proline at position 143 to leucine and alanine at position 284 to glutamic acid. This mutant was then efficiently expressed in Pichia pastoris GS115, and the immobilization conditions were optimized to improve catalytic activity and stability.

Benefits of technology

It significantly improved catalytic activity by 12.5 times, achieved protein expression levels of 1.8~1.9 g/L, reduced production costs, and achieved a conversion rate of 95.6% for vitamin C fatty acid esters and 97.3% for vitamin A oleate in non-aqueous phase reactions, meeting the synthesis requirements of high value-added products.

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Abstract

The invention relates to the technical field of genetic engineering and bioengineering, in particular to a Candida antarctica lipase B mutant and application thereof. The Candida antarctica lipase B mutant provided by the invention is obtained by combined mutation of the 143rd site and the 284th site on an amino acid sequence shown by wild type lipase B. The Candida antarctica lipase B mutant provided by the invention is a Candida antarctica lipase B mutant. Compared with a wild type enzyme, the enzyme activity of the constructed Candida antarctica lipase B mutant is improved by 12.5 times, the protein expression quantity is as high as 1.8-1.9 g / L under the condition of a 5L fermentation tank scale, the yield can be remarkably improved, and the production cost can be reduced. In addition, the Candida antarctica lipase B mutant can be used for synthesis of non-natural substrates with high added values, such as vitamin C fatty acid ester and vitamin A oleate, and lays a foundation for application of lipase in the field of fine chemical engineering.
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