一种基于通道结构入口理性设计提高脂肪酶活性的分子改造方法及其突变体
By modifying key residues at the Tunnel entry point of lipase through molecular simulation and site-directed mutagenesis, the hydrolytic activity of lipase was improved, solving the problem of insufficient catalytic activity for long-chain substrates, and enabling efficient application in the food, pharmaceutical and biochemical fields.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- NANJING TECH UNIV
- Filing Date
- 2025-08-22
- Publication Date
- 2026-07-17
AI Technical Summary
Existing lipases have low catalytic activity for long-chain or large-volume substrates, which limits their effectiveness in industrial applications, especially when preparing drug intermediates with long fatty chains or large steric hindrance.
By identifying key residues at the lipase tunnel entry point through molecular simulation, site-directed saturation mutagenesis was performed using the NNK degenerate codon to establish a mutant library, and the hydrolytic activity of the lipase was improved by microplate screening.
It significantly improved the hydrolytic activity of lipases. Multiple mutants showed more than 20-fold increased hydrolytic activity of pNPP at 40℃, enhancing its industrial application performance in the food, pharmaceutical and biochemical fields.
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Figure CN121075406B_ABST