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Using conventional optimization techniques, physical parameters in submerged fermentation were adjusted at the shake flask level to reach 184 U\/mL. These physicochemical parameters were further optimized by statistical surface response methodology using Box Behnken design, and the protease yield increased to 295 U\/mL. The protease was purified and characterized biochemically. Both Ca<jats:sup>2+<\/jats:sup> and Fe<jats:sup>2+<\/jats:sup> increased the activity of the 36\u00a0kDa protease enzyme. Based on its strong inhibition by ethylenediaminetetracetate (EDTA), the enzyme was confirmed to be a metalloprotease. The protease was also resistant to various organic solvents (benzene, ethanol, methanol), surfactants (Triton X-100), sodium dodecyl sulfate (SDS), Tween 20, Tween-80 and oxidants hydrogen per oxide (H<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub>). Characteristics, such as tolerance to high SDS and H<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub> concentrations, indicate that this protease has potential applications in the pharmaceutical and detergent industries.<\/jats:p>","DOI":"10.1186\/s12896-024-00870-5","type":"journal-article","created":{"date-parts":[[2024,7,15]],"date-time":"2024-07-15T09:02:37Z","timestamp":1721034157000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":22,"title":["Characterization of a thermostable protease from Bacillus subtilis BSP strain"],"prefix":"10.1186","volume":"24","author":[{"given":"Tanveer","family":"Majeed","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charles C.","family":"Lee","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William J.","family":"Orts","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Romana","family":"Tabassum","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tawaf Ali","family":"Shah","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yousef A. 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