{"paper_id":"1a37a04c-7248-4c66-bbfd-bd52fc9350c0","body_text":"Isolation and characterization of Lysobacter Enzymogene PS15 for Protease Production: Optimization Using Response Surface Methodology | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Isolation and characterization of Lysobacter Enzymogene PS15 for Protease Production: Optimization Using Response Surface Methodology Plabon Islam Turzo, Israt Jahan, Aicha Akouz, Sharmin Zaman Emon, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8585128/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract In this study, a novel protease-producing strain, Lysobacter enzymogenes PS15, was successfully isolated from soil and identified through biochemical characterization and 16S rRNA gene sequencing. A sequential optimization strategy was employed to enhance protease yield. The one-factor-at-a-time (OFAT) method first determined an effective nutrient composition of 1.5% dextrose, 1.25% peptone, and 0.6% NaCl, and served as a screening tool to determine the suitable operational ranges for pH (6-8), temperature (34-46 °C), incubation time (12-60 h), and inoculum volume (0.25-3.25 %). These ranges were subsequently optimized using response surface methodology (RSM) using a central composite rotatable design (CCRD). The model predicted a maximum protease activity of 36.62 U mL -1 , which was experimentally confirmed at 35.09 U mL -1 under the optimal conditions (pH 7.0, 39 °C, 47 hours, and 2.3% inoculum volume). The crude enzyme exhibited optimal performance at 40 °C and pH 8, maintaining stability up to 45 °C. This work represents the first RSM-based optimization of protease production from L. enzymogenes , establishing its strong potential for diverse applications, including proteinaceous waste degradation and biocatalysis. Environmental Engineering Applied & Industrial Microbiology General Biochemistry Biotechnology and Bioengineering Enzyme Isolation Lysobacter Protease RSM. Full Text Additional Declarations The authors declare no competing interests. Supplementary Files Supplimentaryfile.docx Supplimentary file floatimage1.jpeg Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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A sequential optimization strategy was employed to enhance protease yield. The one-factor-at-a-time (OFAT) method first determined an effective nutrient composition of 1.5% dextrose, 1.25% peptone, and 0.6% NaCl, and served as a screening tool to determine the suitable operational ranges for pH (6-8), temperature (34-46 °C), incubation time (12-60 h), and inoculum volume (0.25-3.25 %). These ranges were subsequently optimized using response surface methodology (RSM) using a central composite rotatable design (CCRD). The model predicted a maximum protease activity of 36.62 U mL\\u003csup\\u003e-1\\u003c/sup\\u003e, which was experimentally confirmed at 35.09 U mL\\u003csup\\u003e-1 \\u003c/sup\\u003eunder the optimal conditions (pH 7.0, 39 °C, 47 hours, and 2.3% inoculum volume). The crude enzyme exhibited optimal performance at 40 °C and pH 8, maintaining stability up to 45 °C. This work represents the first RSM-based optimization of protease production from \\u003cem\\u003eL. enzymogenes\\u003c/em\\u003e, establishing its strong potential for diverse applications, including proteinaceous waste degradation and biocatalysis.\\u003c/p\\u003e\",\"manuscriptTitle\":\"Isolation and characterization of Lysobacter Enzymogene PS15 for Protease Production: Optimization Using Response Surface Methodology\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2026-01-16 13:12:17\",\"doi\":\"10.21203/rs.3.rs-8585128/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"researchsquare\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":true,\"externalIdentity\":\"\",\"sideBox\":\"\",\"snPcode\":\"\",\"submissionUrl\":\"/submission\",\"title\":\"Research Square\",\"twitterHandle\":\"researchsquare\",\"acdcEnabled\":true,\"dfaEnabled\":false,\"editorialSystem\":\"\",\"reportingPortfolio\":\"\",\"inReviewEnabled\":false,\"inReviewRevisionsEnabled\":true}}],\"origin\":\"\",\"ownerIdentity\":\"326ad82c-8774-4908-9ee2-10fdd4adb96d\",\"owner\":[],\"postedDate\":\"January 16th, 2026\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"posted\",\"subjectAreas\":[{\"id\":61023088,\"name\":\"Environmental Engineering\"},{\"id\":61023089,\"name\":\"Applied \\u0026 Industrial Microbiology\"},{\"id\":61023090,\"name\":\"General Biochemistry\"},{\"id\":61023091,\"name\":\"Biotechnology and Bioengineering\"}],\"tags\":[],\"updatedAt\":\"2026-01-16T13:12:18+00:00\",\"versionOfRecord\":[],\"versionCreatedAt\":\"2026-01-16 13:12:17\",\"video\":\"\",\"vorDoi\":\"\",\"vorDoiUrl\":\"\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-8585128\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-8585128\",\"identity\":\"rs-8585128\",\"version\":[\"v1\"]},\"buildId\":\"XKTyCvWXoU3ODBz1xrDgd\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}