Construction of R16F and D19L mutations in the loop I of bile salt hydrolase (BSH) enzyme from Lactobacillus plantarum B14 and structural and functional analysis of the mutant BSHs

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dc.authorid0000-0001-6906-5257en_US
dc.authorid0000-0003-4367-1747
dc.authorid0000-0003-2701-3250
dc.authorid0000-0003-2928-2577
dc.contributor.authorÖztürk, Mehmet
dc.contributor.authorHacıbeyoğlu, Kübra
dc.contributor.authorÖnal, Cansu
dc.contributor.authorKılıçsaymaz, Zekiye
dc.date.accessioned2021-06-23T19:51:50Z
dc.date.available2021-06-23T19:51:50Z
dc.date.issued2019
dc.departmentBAİBÜ, Fen Edebiyat Fakültesi, Biyoloji Bölümüen_US
dc.description.abstractBile salt hydrolase (BSH), found commonly in intestinal species of Lactobacillus and Bifidobacterium, catalyzes the hydrolysis of glycine or taurine-conjugated bile acids into the amino acids and free bile acids. Deconjugated bile acids potentially play an important role in the reduction of blood cholesterol level and formation of some gastrointestinal diseases such as cholestasis, gallstone formation, and colon cancer. Although the crystal and three-dimensional structures of BSH enzyme are known, the working mechanism of catalytic activity of such an important BSH enzyme is not known very well. Previous in silico analysis of multiple BSH has identified that Arginine-16 (R16) and Aspartate-19 (D19) were catalytically important residues in the active site of BSH. To confirm the function of these amino acids, in this study, BSH enzyme from Lactobacillus plantarum B14 strain was cloned into Escherichia coli and strictly conserved polar R16 and D19 amino acids of BSH enzyme were substituted for hydrophobic Phenylalanine-16 (F16) and Leucine-19 (L19) amino acids, respectively, by polymerase chain reaction (PCR)-based site-directed mutagenesis. The effects of the mutations on catalytic activity and structure of the BSH enzymes were detected by ninhidrin assay and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis, respectively. Research results showed that although F16 mutation led to loss of enzyme activity completely, L19 mutation led to abolishment of the synthesis of BSH enzyme. These results indicated that R16 and D19 amino acids located in loop I of the BSH enzyme might be critical for catalytic activity and assembly of the BSH enzyme respectively.en_US
dc.identifier.doi10.1080/08905436.2019.1570853
dc.identifier.endpage141en_US
dc.identifier.issn0890-5436
dc.identifier.issn1532-4249
dc.identifier.issue2en_US
dc.identifier.scopus2-s2.0-85063262246en_US
dc.identifier.scopusqualityQ2en_US
dc.identifier.startpage125en_US
dc.identifier.urihttps://doi.org/10.1080/08905436.2019.1570853
dc.identifier.urihttps://hdl.handle.net/20.500.12491/10056
dc.identifier.volume33en_US
dc.identifier.wosWOS:000462093900002en_US
dc.identifier.wosqualityQ4en_US
dc.indekslendigikaynakWeb of Scienceen_US
dc.indekslendigikaynakScopusen_US
dc.institutionauthorÖztürk, Mehmet
dc.institutionauthorHacıbeyoğlu, Kübra
dc.institutionauthorÖnal, Cansu
dc.institutionauthorKılıçsaymaz, Zekiye
dc.language.isoenen_US
dc.publisherTaylor & Francis Incen_US
dc.relation.ispartofFood Biotechnologyen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/closedAccessen_US
dc.subjectBile Salt Hydrolaseen_US
dc.subjectCatalytic Activityen_US
dc.subjectLactobacillus Plantarumen_US
dc.subjectProbioticsen_US
dc.subjectSubstrate Preferenceen_US
dc.titleConstruction of R16F and D19L mutations in the loop I of bile salt hydrolase (BSH) enzyme from Lactobacillus plantarum B14 and structural and functional analysis of the mutant BSHsen_US
dc.typeArticleen_US

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