Tanveer Majeed, Charles C Lee, William J Orts, Romana Tabassum, Tawaf Ali Shah, Yousef A Bin Jardan, Turki M Dawoud, Mohammed Bourhia
{"title":"枯草芽孢杆菌 BSP 菌株的一种恒温蛋白酶的特性。","authors":"Tanveer Majeed, Charles C Lee, William J Orts, Romana Tabassum, Tawaf Ali Shah, Yousef A Bin Jardan, Turki M Dawoud, Mohammed Bourhia","doi":"10.1186/s12896-024-00870-5","DOIUrl":null,"url":null,"abstract":"<p><p>This study used conservative one variable-at-a-time study and statistical surface response methods to increase the yields of an extracellular thermostable protease secreted by a newly identified thermophilic Bacillus subtilis BSP strain. 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<sup>2+</sup> and Fe<sup>2+</sup> increased the activity of the 36 kDa 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<sub>2</sub>O<sub>2</sub>). Characteristics, such as tolerance to high SDS and H<sub>2</sub>O<sub>2</sub> concentrations, indicate that this protease has potential applications in the pharmaceutical and detergent industries.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"24 1","pages":"49"},"PeriodicalIF":3.5000,"publicationDate":"2024-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11247832/pdf/","citationCount":"0","resultStr":"{\"title\":\"Characterization of a thermostable protease from Bacillus subtilis BSP strain.\",\"authors\":\"Tanveer Majeed, Charles C Lee, William J Orts, Romana Tabassum, Tawaf Ali Shah, Yousef A Bin Jardan, Turki M Dawoud, Mohammed Bourhia\",\"doi\":\"10.1186/s12896-024-00870-5\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>This study used conservative one variable-at-a-time study and statistical surface response methods to increase the yields of an extracellular thermostable protease secreted by a newly identified thermophilic Bacillus subtilis BSP strain. 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<sup>2+</sup> and Fe<sup>2+</sup> increased the activity of the 36 kDa 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<sub>2</sub>O<sub>2</sub>). Characteristics, such as tolerance to high SDS and H<sub>2</sub>O<sub>2</sub> concentrations, indicate that this protease has potential applications in the pharmaceutical and detergent industries.</p>\",\"PeriodicalId\":8905,\"journal\":{\"name\":\"BMC Biotechnology\",\"volume\":\"24 1\",\"pages\":\"49\"},\"PeriodicalIF\":3.5000,\"publicationDate\":\"2024-07-15\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11247832/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"BMC Biotechnology\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://doi.org/10.1186/s12896-024-00870-5\",\"RegionNum\":3,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"BIOTECHNOLOGY & APPLIED MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"BMC Biotechnology","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1186/s12896-024-00870-5","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
Characterization of a thermostable protease from Bacillus subtilis BSP strain.
This study used conservative one variable-at-a-time study and statistical surface response methods to increase the yields of an extracellular thermostable protease secreted by a newly identified thermophilic Bacillus subtilis BSP strain. 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 Ca2+ and Fe2+ increased the activity of the 36 kDa 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 (H2O2). Characteristics, such as tolerance to high SDS and H2O2 concentrations, indicate that this protease has potential applications in the pharmaceutical and detergent industries.
期刊介绍:
BMC Biotechnology is an open access, peer-reviewed journal that considers articles on the manipulation of biological macromolecules or organisms for use in experimental procedures, cellular and tissue engineering or in the pharmaceutical, agricultural biotechnology and allied industries.