分子改良提高甘露聚糖酶的耐热性  

Improving the thermotolerance of mannanase through molecular improvement

作　　者：王亚伟 薛强 熊海容 WANG Ya-wei;XUE Qiang;XIONG Hai-rong(School of Life Science and Technology,Wuhan Polytechnic University,Wuhan 430048,China;College of Life Sciences,South-Central Minzu University,Wuhan 430074,China)

机构地区：[1]武汉轻工大学生命科学与技术学院,武汉430048 [2]中南民族大学生命科学学院,武汉430074

出　　处：《湖北农业科学》2023年第12期151-153,162,共4页Hubei Agricultural Sciences

基　　金：湖北省技术创新重大项目(2018ABA093)。

摘　　要：以来源于嗜热放线菌(Thermobifida fusca)的甘露聚糖酶基因(KJ806638)为研究对象,利用易错PCR方法对其进行改造,获得了耐热性进一步提高的甘露聚糖酶。筛选获得的N16I突变株,其最适反应温度从72.5℃提升到77.5℃。差示扫描荧光定量(DSF)检测结果显示,酶蛋白质的表观解链温度Tm从61.9℃提升到63.2℃。The mannanase gene(KJ806638)from Thermobifida fusca was used as the research object,and it was modified by the er⁃ror-prone PCR method,and the heat resistance of mannanase was further improved.The optimum reaction temperature of N16I mutant was increased from 72.5℃to 77.5℃.Differential scanning fluorometry(DSF)testing results showed that the apparent decomposition temperature Tm of the enzyme protein increased from 61.9℃to 63.2℃.

关 键 词：甘露聚糖酶 易错PCR 热稳定性 

分 类 号：Q55[生物学—生物化学]