Abstract

Organic solvent-tolerant proteases have many applications in the synthesis of peptides. In this study, we have developed a low-cost and convenient method to produce highly concentrated organic solvent-tolerant protease. Organic solvent tolerant protease (OSP) gene from Bacillus sphaericus DS11 was cloned and expressed in Bacillus subtilis WB800. The optimum pH of the recombinant protease was 9.0. The optimum temperature of the recombinant protease was 40 °C. The recombinant protease was purified by ethanol with the yield of (87.33%). The yield of OSP enriched by ethanol was higher than that of by Ni-chelating affinity chromatography, which indicated that precipitation of the recombinant OSP with ethanol is a relatively low-cost and fast method for organic solvent -tolerant protease preparation. These results showed that this enzyme could be very useful in different industrial applications.

摘要

耐有机溶剂的蛋白酶在肽合成中有许多应用。在这项研究中，我们已经开发了一种低成本，便捷的方法来生产高浓度的耐有机溶剂的蛋白酶。克隆了球形芽孢杆菌DS11的有机溶剂耐受蛋白酶（OSP）基因并在枯草芽孢杆菌中表达WB800。重组蛋白酶的最佳pH为9.0。重组蛋白酶的最佳温度为40°C。用乙醇纯化重组蛋白酶，产率为（87.33％）。乙醇富集的OSP的产率高于Ni-螯合亲和色谱法的产率，这表明用乙醇沉淀重组OSP是一种相对低成本，快速的耐有机溶剂蛋白酶的制备方法。这些结果表明该酶在不同的工业应用中可能非常有用。