@article { author = {Mohamadi, Shohreh and Mehrabi, Maryam and Sajadimajd, Soraya}, title = {Purification and Characterization of an Extracellular Alkaline Solvent-stable Metalloprotease Secreted from Newly Isolated Bacillus sp. DEM05: Optimization of Protease Production}, journal = {Iranian Journal of Biotechnology}, volume = {19}, number = {3}, pages = {28-37}, year = {2021}, publisher = {National Institute of Genetic Engineering and Biotechnology of Iran}, issn = {1728-3043}, eissn = {2322-2921}, doi = {10.30498/ijb.2021.247161.2866}, abstract = {Background: Proteases play an important role in food, leather, detergent, and medical technologies.Objectives: In the current study, an alkaliphilic solvent-stable thermotolerant metalloprotease was isolated from Bacillus sp. DEM05.Material and Methods: For culture optimization, carbon, and nitrogen sources as well as incubation temperature, pH, and time were examined.Results: Herein the highest outcome for bacterial growth and protease production was obtained after 72 h incubation (pH 7) at 37 °C. DEM05 protease was successfully purified and the specific activity of the protease was 1075 U.mg-1. The purity of the enzyme was verified by SDS-PAGE electrophoresis as a single band of 30 kDa. The optimal activity of the enzyme was at pH 10 and 50 °C. H2O2, SDS, Triton X-100, Zn2+, Co2+, and Cu2+ could increase the protease activity. EDTA inhibited the protease activity, revealed that it can be classified as a metalloprotease. The enzyme was compatible with the water-miscible and water-immiscible organic solvents and proteolyzed several substrates, implying the wide substrate specificity.Conclusions: The results brought convincing evidence that DEM05 protease could be recruited as a novel prevailing protease that can be earmarked on industrial and medical technologies.}, keywords = {Alkaline,Bacillus sp,optimization,Protease,Purification}, url = {https://www.ijbiotech.com/article_135047.html}, eprint = {https://www.ijbiotech.com/article_135047_44dea876d1b451ce8d15248531282734.pdf} }