Production, purification and characterisation of thermostable metallo-protease from newly isolated bacillus sp KG5

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dc.contributor.authorAhmetoğlu, Nazenin
dc.contributor.authorBekler, Fatma Matpan
dc.contributor.authorAcer, Ömer
dc.contributor.authorGüven, Reyhan Gül
dc.contributor.authorGüven, Kemal
dc.date.accessioned2024-04-24T17:58:21Z
dc.date.available2024-04-24T17:58:21Z
dc.date.issued2015
dc.departmentDicle Üniversitesi, Fen Fakültesi, Moleküler Biyoloji ve Gentik Bölümüen_US
dc.description.abstractBackground: Due to the importance of microbial proteases in biotechnological applications, a number of microorganisms are being explored. The production, purification and characterisation of extracellular metallo-proteases by producing Bacillus sp. KG5 was studied. Material and Methods: Bacterial strain KG5 was isolated from Kös (Bingöl) hot spring. The strain KG5 was identified by morphological, physiological, biochemical and 16S rRNA gene sequencing. The effects of various parameters on protease production, such as time, temperature, pH, carbon and nitrogen sources and CaCl2were studied. The enzyme was purified by ammonium sulphate precipitation and Sephadex G-75 gel permeation chromatography. Molecular weight was calculated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and zymographic analysis. The effects of some metal ions, chelators and inhibitors on enzyme activity were determined. Results: The optimum temperature, pH and incubation period for protease production were 40-45°C, 7.0 and 24 h, respectively. It was determined that the best nitrogen sources were yeast extract and urea, while the best carbon sources were lactose and galactose. However, glucose as a source of carbon was found to inhibit the production of the enzyme. The maximum enzyme production was increased in the presence of CaCl2. The molecular weight of purified enzyme was found to be approximately 48 kDa. It was found that the enzyme was fully stable in the presence of 2 mM CaCl2 at 50°C after 120 min. Purified protease was significantly activated by Ca2+ and Mg2+, while it was greatly inhibited by Cu2+, Zn2+, Hg2+ and SDS as well as by the metal ion chelators ethylenediaminetetraacetic (EDTA) and 1,10-phenanthroline. Phenylmethylsulfonyl fluoride (PMSF) had a little effect on the enzyme. Conclusions: Our findings suggest the potential of this isolate for protease production and that this enzyme may be suitable for biotechnological applications.en_US
dc.identifier.citationAhmetoğlu, N., Bekler, F. M., Acer, Ö., Güven, R. G. ve Güven, K. (2015). Production, purification and characterisation of thermostable metallo-protease from newly isolated bacillus sp KG5. EurAsian Journal of BioSciences, 9, 1-11.
dc.identifier.doi10.5053/ejobios.2015.9.0.1
dc.identifier.endpage11en_US
dc.identifier.issn1307-9867
dc.identifier.scopus2-s2.0-84925882556
dc.identifier.scopusqualityN/A
dc.identifier.startpage1en_US
dc.identifier.urihttps://doi.org/10.5053/ejobios.2015.9.0.1
dc.identifier.urihttps://hdl.handle.net/11468/23854
dc.identifier.volume9en_US
dc.indekslendigikaynakScopus
dc.language.isoenen_US
dc.publisherFoundation for Enviromental Protection and Researchen_US
dc.relation.ispartofEurAsian Journal of BioSciences
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/closedAccessen_US
dc.subjectBacillus Sp. Kg5en_US
dc.subjectBiotechnologyen_US
dc.subjectProtease production and characterisationen_US
dc.titleProduction, purification and characterisation of thermostable metallo-protease from newly isolated bacillus sp KG5en_US
dc.titleProduction, purification and characterisation of thermostable metallo-protease from newly isolated bacillus sp KG5
dc.typeArticleen_US

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