Expression and characterization of a novel organic solvent tolerant protease from Bacillus sphaericus DS11

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.

Keywords:

• Bacillus sphaericus
• heterologous expression
• oligopeptides synthesis
• organic solvent-tolerant protease

Disclosure statement

The authors declare no conflict of interest.

Additional information

Funding

This work was supported by the National Natural Science Foundation of China [grant no. 31772016], Jiangsu Province Marine Science and Technology Innovation Project [HY2018-10], the Priority Academic Program Development of Jiangsu Higher Education Institutions, Six Talent Peaks Project in Jiangsu Province [2016-SWYY-195], and Project “333” of Jiangsu Province, Open-end Funds of Jiangsu Key Laboratory of Marine Bioresources and Environment 296 [SH20191204], and “521 high-level talents training project” in Lianyungang.