{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T19:48:04Z","timestamp":1789501684552,"version":"build-2803163510"},"reference-count":106,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2021,1,18]],"date-time":"2021-01-18T00:00:00Z","timestamp":1610928000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>A novel temperature stable alkaline protease yielding bacteria was isolated from the soils of Dachigam National Park, which is known to be inhabited by a wide variety of endemic plant and animal species of Western Himalaya. This high-potential protease producing isolate was characterized and identified as Bacillus amyloliquefaciens strain HM48 by morphological, Gram\u2019s staining and biochemical techniques followed by molecular characterization using 16S rRNA approach. The extracellular protease of B. amyloliquefaciens HM48 was purified by precipitating with ammonium sulfate (80%), followed by dialysis and Gel filtration chromatography increasing its purity by 5.8-fold. The SDS\u2013PAGE analysis of the purified enzyme confirmed a molecular weight of about \u224825 kDa. The enzyme displayed exceptional activity in a broad temperature range (10\u201390 \u00b0C) at pH 8.0, retaining its maximum at 70 \u00b0C, being the highest reported for this proteolytic Bacillus sp., with KM and Vmax of 11.71 mg\/mL and 357.14 \u00b5mol\/mL\/min, respectively. The enzyme exhibited remarkable activity and stability against various metal ions, surfactants, oxidizing agent (H2O2), organic solvents and displayed outstanding compatibility with widely used detergents. This protease showed effective wash performance by exemplifying complete blood and egg-yolk stains removal at 70 \u00b0C and efficiently disintegrated chicken feathers making it of vital importance for laundry purpose and waste management. For functional analysis, protease gene amplification of strain HM48 yielded a nucleotide sequence of about 700 bp, which, when checked against the available sequences in NCBI, displayed similarity with subtilisin-like serine protease of B. amyloliquefaciens. The structure of this protease and its highest-priority substrate \u03b2-casein was generated through protein modeling. These protein models were validated through futuristic algorithms following which protein\u2013protein (protease from HM48 and \u03b2-casein) docking was performed. The interaction profile of these proteins in the docked state with each other was also generated, shedding light on their finer details. Such attributes make this thermally stable protease novel and suitable for high-temperature industrial and environmental applications.<\/jats:p>","DOI":"10.3390\/biom11010117","type":"journal-article","created":{"date-parts":[[2021,1,19]],"date-time":"2021-01-19T04:55:31Z","timestamp":1611032131000},"page":"117","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":29,"title":["Biochemical Characterization and Functional Analysis of Heat Stable High Potential Protease of Bacillus amyloliquefaciens Strain HM48 from Soils of Dachigam National Park in Kashmir Himalaya"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3549-2546","authenticated-orcid":false,"given":"Hina","family":"Mushtaq","sequence":"first","affiliation":[{"name":"Department of Environmental Science, University of Kashmir, Srinagar, Jammu &amp; Kashmir 190006, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arshid","family":"Jehangir","sequence":"additional","affiliation":[{"name":"Department of Environmental Science, University of Kashmir, Srinagar, Jammu &amp; Kashmir 190006, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shabir Ahmad","family":"Ganai","sequence":"additional","affiliation":[{"name":"Division of Basic Sciences and Humanities, Faculty of Agriculture, SKUAST-Kashmir, Sopore 193201, Jammu &amp; Kashmir, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5226-6212","authenticated-orcid":false,"given":"Saleem","family":"Farooq","sequence":"additional","affiliation":[{"name":"Department of Environmental Science, University of Kashmir, Srinagar, Jammu &amp; Kashmir 190006, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bashir Ahmad","family":"Ganai","sequence":"additional","affiliation":[{"name":"Centre of Research for Development, University of Kashmir, Srinagar, Jammu &amp; Kashmir 190006, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruqeya","family":"Nazir","sequence":"additional","affiliation":[{"name":"Centre of Research for Development, University of Kashmir, Srinagar, Jammu &amp; Kashmir 190006, India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,1,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"137","DOI":"10.5958\/2320-642X.2015.00015.0","article-title":"Phytodiversity and Seasonal Variations in the Soil Characteristics of Shrublands of Dachigam National Park, Jammu and Kashmir, India","volume":"3","author":"Yaqoob","year":"2015","journal-title":"Clim. 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