分子改良提高t露聚糖酶的耐热性

doi:10.14088/j.cnki.issn0439-8114.2023.12.027

湖北农业科学 ›› 2023, Vol. 62 ›› Issue (12): 151-153,162.doi: 10.14088/j.cnki.issn0439-8114.2023.12.027

分子改良提高t露聚糖酶的耐热性

王亚伟¹, 薛强², 熊海容²

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

收稿日期:2023-09-25 发布日期:2024-01-10
通讯作者: 熊海容,教授,主要从事酶蛋白质工程研究,（电子信箱）995025147@qq.com。
作者简介:王亚伟,讲师,博士,主要从事微生物学研究,（电子信箱）253182556@qq.com.
基金资助:
湖北省技术创新重大项目（2018ABA093）

Improving the thermotolerance of mannanase through molecular improvement

WANG Ya-wei¹, XUE Qiang², XIONG Hai-rong²

1. School of Life Science and Technology,Wuhan Polytechnic University,Wuhan 430048,China;
2. College of Life Sciences, South-Central Minzu University, Wuhan 430074,China

Received:2023-09-25 Published:2024-01-10

摘要/Abstract

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T_m从61.9 ℃提升到 63.2 ℃。

关键词: 甘露聚糖酶, 易错PCR, 热稳定性

Abstract: The mannanase gene （KJ806638） from Thermobifida fusca was used as the research object, and it was modified by the error-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 T_m of the enzyme protein increased from 61.9 ℃ to 63.2 ℃.

Key words: mannanase, error-prone PCR, thermostabilty

中图分类号:

王亚伟, 薛强, 熊海容. 分子改良提高t露聚糖酶的耐热性[J]. 湖北农业科学, 2023, 62(12): 151-153,162.

WANG Ya-wei, XUE Qiang, XIONG Hai-rong. Improving the thermotolerance of mannanase through molecular improvement[J]. HUBEI AGRICULTURAL SCIENCES, 2023, 62(12): 151-153,162.

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分子改良提高t露聚糖酶的耐热性

Improving the thermotolerance of mannanase through molecular improvement

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