耐热碱性蛋白酶AH-101的异源表达、酶学性质与洗涤应用研究
Heterologous Expression,Characterization and Washing Application of Thermostable Alkaline Protease AH-101
郝漫 1邵岚莹 1公为峰 1惠威 1史超硕 1路福平 1张会图1
作者信息
- 1. 天津科技大学生物工程学院 天津 300457
- 折叠
摘要
目的:通过NCBI(National Center for Biotechnology Information)数据库筛选可以应用于洗涤工业的蛋白酶,使用地衣芽孢杆菌进行高效表达.方法:利用同源重组的方式将蛋白酶基因整合到地衣芽孢杆菌基因组中,研究重组蛋白酶的酶学特性和洗涤效果.结果:该重组酶的最适作用温度为60℃,最适pH为11.0;AH-101具有良好的耐热性,可以在20～55℃范围内保持稳定,在60℃保温1 h,剩余活性仍高于30％;此外,该酶与表面活性剂和液体洗涤剂具有良好的兼容性,在终浓度为0.1％和0.5％的表面活性剂和液体洗涤剂中孵育2 h,重组酶的酶活力损失小于30％.结论:成功构建了一株含有外源蛋白酶的地衣芽孢杆菌重组菌株,对重组蛋白酶的特性和洗涤效果进行研究,该重组蛋白酶可以作为环保型酶制剂应用到洗涤工业中.
Abstract
Objective:To screen the protease which can be used in washing industry through NCBI database and express it efficiently using Bacillus licheniformis.Methods:The protease gene was integrated into the genome of Bacillus licheniformis by homologous recombination,the fermentation was studied,and the enzymatic characteristics and washing effect of the recombinant protease were investigated.Results:The optimum temperature and pH of the recombinant enzyme were 60℃ and 11.0,respectively.AH-101 has good heat resistance,which can be kept stable in the range of 20～55℃,and the residual activity is still higher than 30％at 60℃ for 1 h.In addition,the enzyme has good compatibility with surfactants and liquid detergents,and the loss of activity of the recombinant enzyme is less than 30％when incubated for 2 h in surfactants and liquid detergents with final concentrations of 0.1％and 0.5％.Conclusion:A recombinant Bacillus licheniformis strain containing exogenous protease was successfully constructed,and the properties and washing effect of the recombinant protease were investigated.The recombinant protease can be used as an environmentally-friendly enzyme preparation in the washing industry.
关键词
耐热碱性蛋白酶/地衣芽孢杆菌/整合表达/洗涤工业Key words
Thermostable alkaline protease/Bacillus licheniformis/Integrated/expression/Washing application引用本文复制引用
基金项目
国家重点研发计划(2021YFC2100400)
出版年
2024
NETL
NSTL
万方数据