{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,6]],"date-time":"2026-05-06T08:49:14Z","timestamp":1778057354563,"version":"3.51.4"},"reference-count":30,"publisher":"Bentham Science Publishers Ltd.","issue":"9","content-domain":{"domain":["eurekaselect.com"],"crossmark-restriction":true},"short-container-title":["PPL"],"published-print":{"date-parts":[[2023,9]]},"abstract":"<jats:sec>\n<jats:title>Background:<\/jats:title>\n<jats:p>An increasing prevalence of biofilm forming strains by vancomycinresistance\nStaphylococcus aureus (VRSA) is one of the most important causes of antimicrobial\nresistance. VRSA possesses various regulatory factors to form and sustain biofilm in biotic or abiotic\nconditions. Among them, ureolytic activity is an important factor in the stabilization of biofilms by\nneutralizing the acidic environment. Various urease accessory proteins are required to activate the\nurease enzyme inside the biofilm.<\/jats:p>\n<\/jats:sec>\n<jats:sec>\n<jats:title>Objective:<\/jats:title>\n<jats:p>To optimize the cloning, expression and purification of urease accessory protein E from\nVRSA for determination of the secondary structure, and functional characterization by using\nBerthelot\u2019s method.<\/jats:p>\n<\/jats:sec>\n<jats:sec>\n<jats:title>Methods:<\/jats:title>\n<jats:p>BAB58453.1 gene (which encodes possible urease accessory protein E), having 38%\nsimilarity to Bacillus pasteurii UreE protein, was cloned, expressed, and purified by single-step\naffinity chromatography for performing secondary structural studies using circular dichroism\nspectroscopy, and functional analysis using Berthelot\u2019s and crystal violet assay.<\/jats:p>\n<\/jats:sec>\n<jats:sec>\n<jats:title>Results:<\/jats:title>\n<jats:p>Structure elucidation using NMR and circular dichroism spectroscopy techniques revealed\nthat UreE protein has a partially folded\u03b1-helical structure. Using Berthelot's method, it was identified\nthat the purified UreE protein has enhanced urease enzyme activity, in comparison to the control.\nFrom the results of Berthelot\u2019s and crystal violet assays, it was deduced that the selected gene (UreE\nprotein) plays a key role in enhancing urease enzyme activity and contributes to biofilm stability.<\/jats:p>\n<\/jats:sec>\n<jats:sec>\n<jats:title>Conclusion:<\/jats:title>\n<jats:p>Structural studies on VRSA urease accessory proteins could aid in the identification of\nnew drug targets or the development of effective antibiofilm strategies (in combination with other drug\ntargets) against infections caused by biofilm-producing strains.<\/jats:p>\n<\/jats:sec>","DOI":"10.2174\/0929866530666230801163340","type":"journal-article","created":{"date-parts":[[2023,8,3]],"date-time":"2023-08-03T05:12:09Z","timestamp":1691039529000},"page":"754-762","update-policy":"https:\/\/doi.org\/10.2174\/bsp_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Structural and Functional Analysis of Urease Accessory Protein E from\nVancomycin-Resistance Staphylococcus aureus MU50 Strain"],"prefix":"10.2174","volume":"30","author":[{"given":"Humaira","family":"Siddiqui","sequence":"first","affiliation":[{"name":"Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological\nSciences, University of Karachi, Karachi-75270, Pakistan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2730-5547","authenticated-orcid":true,"family":"Atia-tul-Wahab","sequence":"additional","affiliation":[{"name":"Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological\nSciences, University of Karachi, Karachi-75270, Pakistan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aftab","family":"Ahmed","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Chapman University, Irvine, CA\n92618, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M. 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