Purification and characterization of glucoamylase from Bacillus sp. isolated from root flora of Prunus mahaleb tree by LC-MS/MS analysis

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dc.contributor.authorÇelik, Muhammed Safa
dc.contributor.authorKapancık, Serkan
dc.contributor.authorSeyran, Esra
dc.contributor.authorÇetinkaya, Serap
dc.date.accessioned2025-05-04T16:42:05Z
dc.date.available2025-05-04T16:42:05Z
dc.date.issued2025
dc.departmentSivas Cumhuriyet Üniversitesi
dc.description.abstractGlucoamylases are enzymes that release free glucose by hydrolyzing consecutive α-1,4 bonds at the non-reducing ends of starch molecules. This enzyme was purified using ammonium sulfate precipitation, and its molecular mass was determined to be approximately 65.2 kDa via SDS-PAGE. Using zymogram analysis, the purified sample's active glucoamylase content was verified. Nano-Liquid Chromatography Mass Spectrometry (nLC-MS/MS) characterization identified peptides covering 46 % of the glucoamylase protein sequence, indicating partial characterization of the enzyme. It was found that the ideal pH and temperature for glucoamylase activity were 6.0 and 37 °C, respectively. Using soluble starch as a substrate, the kinetic parameters were calculated: the Km (substrate concentration at half-maximal velocity) was 30.21 μM, and the Vmax (maximum reaction velocity) was 35.59 μmol mg protein−1 min−1. The enzyme demonstrated optimal activity with soluble starch, highlighting its specificity for starch hydrolysis. Additionally, the enzymatic activity was enhanced in the presence of CaCl2, indicating a positive effect of calcium ions. Its optimal conditions and kinetic parameters provide valuable insights for its industrial and biomedical use. © 2025 Elsevier B.V.
dc.description.sponsorshipScientific Research Projects of Sivas Cumhuriyet University, (F-2022-674)
dc.identifier.doi10.1016/j.ijbiomac.2025.143078
dc.identifier.issn0141-8130
dc.identifier.scopus2-s2.0-105002396030
dc.identifier.scopusqualityQ1
dc.identifier.urihttps://doi.org/10.1016/j.ijbiomac.2025.143078
dc.identifier.urihttps://hdl.handle.net/20.500.12418/35063
dc.identifier.volume309
dc.indekslendigikaynakScopus
dc.language.isoen
dc.publisherElsevier B.V.
dc.relation.ispartofInternational Journal of Biological Macromolecules
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanı
dc.rightsinfo:eu-repo/semantics/closedAccess
dc.snmzKA_Scopus_20250504
dc.subjectBacillus sp.
dc.subjectCharacterization
dc.subjectGlucoamylase
dc.subjectIsolation
dc.subjectLC-MS/MS
dc.subjectPurification
dc.titlePurification and characterization of glucoamylase from Bacillus sp. isolated from root flora of Prunus mahaleb tree by LC-MS/MS analysis
dc.typeArticle

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