疏棉状嗜热丝孢菌(Thermomyces lanuginosus)脂肪酶的理性设计提高其活性和温度稳定性

目的:通过对疏棉状嗜热丝孢菌(Thermomyces lanuginosus)脂肪酶的理性设计,获得高酶活与耐高温的脂肪酶品种,为脂肪酶在饲料、油脂加工和生物柴油等领域的应用奠定基础.方法:对脂肪酶典型结构域lid和loop区域的系统发育分析,找到候选的位点,理性设计并通过实验验证,获得脂肪酶活性和耐高温特性显著提高的...

Improving the Activity and Thermal Stability of Thermomyces lanuginosus Lipase by Rational Design

Objective:In order to obtain a lipase with high enzyme activity and high temperature resistance,the Thermomyces lanuginosus lipase was modified by rational design,which laid the foundation for the application of lipase in the feed industry,oil processing and biodiesel.Methods:A candidate site was found by phylogenetic analysis of the lid and loop regions of the typical domain of lipase.A lipase recombinant with significantly improved lipase activity and high temperature resistance was obtained through rational design and experimental verification,and a multi-copy vector was constructed to complete the evaluation of the enzyme production ability in a 50 L fermentor.Results:The thermal stability of lipase was significantly improved after designing.The crude enzyme solution still retained 78.94%of the enzyme activity after being placed at 80℃for 12 h.After fermentation for 168 h in a 50 L fermentor,the enzyme activity of the supernatant reached 29000 U/m L.Conclusion:A novel high activity and high thermal stable lipase was successfully designed and obtained,which facilitates its large-scale production and industrial application.