ARTP诱变选育高温蛋白酶高产菌株及其酶学性质研究

Selection of high- yield thermostable protease producing strain by ARTP and the study on its enzymological properties

摘要: 以高温环境土样中筛选的产高温蛋白酶菌为出发菌株,利用常压室温等离子体诱变技术选育出一株高产突变株Bacillus licheniformis TP1-5,产酶活力达到33200U/m L,为出发菌株的1.56倍。通过乙醇沉淀和阴离子交换层析两步纯化,获得电泳纯的高温蛋白酶。酶学性质研究表明,该蛋白酶为高温碱性丝氨酸蛋白酶,酶的最适p H为10.0,最适反应温度为60℃,具有较好的p H稳定性和热稳定性;对离子型和非离子型表面活性剂均具有很高的耐受性,为进一步开发应用奠定了基础。

Abstract: In order to obtain an industrial strain with higher thermostable protease production,the original strain of Bacillus licheniformis TP1 was mutated by atmospheric and room temperature plasmas. A good genetic stability strain TP1- 5 was screened from the mutants,which produced 33200 U / m L from initial 21300 U / m L of the thermostable protease.The protease was purified by ethanol precipitation and anion exchange chromatography to electrophoretic homogeneity. The purified protease had a good p H stability and thermal stability,with optimal temperature and p H was 60℃ and 10.0. The protease also exhibited high stability towards surfactants such as SDS,Tween-80,Triton X-100 et al.