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

作　　者：郝漫 邵岚莹 公为峰 惠威 史超硕 路福平[1] 张会图[1] HAO Man;SHAO Lanying;GONG Weifeng;HUI Wei;SHI Chaoshuo;LU Fuping;ZHANG Huitu(College of Biotechnology,Tianjin University of Science&Technology,Tianjin 300457,China)

机构地区：[1]天津科技大学生物工程学院,天津300457

出　　处：《中国生物工程杂志》2024年第4期23-32,共10页China Biotechnology

基　　金：国家重点研发计划(2021YFC2100400)资助项目。

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。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 

分 类 号：Q814[生物学—生物工程]