Cloning, Over-Expression, and Purification of ?–Carbonic Anhydrasefrom an Extremophilic Bacterium: Deinococcus radiodurans

dc.contributor.authorKafa, Ayşe Hümeyra Taşkın
dc.contributor.authorCihan, Arzu Çöleri
dc.contributor.authorKuzucu, Mehmet
dc.contributor.authorÇankaya, Murat
dc.date.accessioned2024-10-26T17:33:45Z
dc.date.available2024-10-26T17:33:45Z
dc.date.issued2021
dc.departmentSivas Cumhuriyet Üniversitesi
dc.description.abstractIn this study, the cloning, purification and initial characterization of carbonic anhydrase (DrCA) enzyme which we consider to be important in the resistance physiology from extremely radioresistant bacteria Deinococcus radiodurans is performed. In addition, the effect of increased gamma irradiation doses on pH-related DrCA enzyme activity was determined. DrCA activity after radiation treatment showed that the activity continuously increased by 6 fold, up to the first 800 Gy, which a decrease in activity was observed thereafter. The maximum $CO_2$ hydration activity for DrCA enzyme was observed at pH 7.0 and 40°C. DrCA enzyme, homo-dimer complex, is slightly thermostable. The activity of DrCA was significantly enhanced by several metal ions, especially $Zn^{2+}$, which resulted in 5-fold increases of $CO_2$ hydration activity. Also sulfonamide showed inhibitory effect on the pure enzyme. The apparent Km and Vmax for $CO_2$ as substrate were 8.4 mM and 637 WAU/mg for DrCA respectively. The $CO_2$ hydration assay demonstrated that the specific activity of purified recombinant enzymes (DrCA) was significantly high.
dc.identifier.doi10.15671/hjbc.775093
dc.identifier.endpage231
dc.identifier.issn1303-5002
dc.identifier.issn2687-475X
dc.identifier.issue3
dc.identifier.startpage219
dc.identifier.trdizinid461860
dc.identifier.urihttps://doi.org/10.15671/hjbc.775093
dc.identifier.urihttps://search.trdizin.gov.tr/tr/yayin/detay/461860
dc.identifier.urihttps://hdl.handle.net/20.500.12418/23283
dc.identifier.volume49
dc.indekslendigikaynakTR-Dizin
dc.language.isoen
dc.relation.ispartofHacettepe Journal of Biology and Chemistry
dc.relation.publicationcategoryMakale - Ulusal Hakemli Dergi - Kurum Öğretim Elemanı
dc.rightsinfo:eu-repo/semantics/openAccess
dc.titleCloning, Over-Expression, and Purification of ?–Carbonic Anhydrasefrom an Extremophilic Bacterium: Deinococcus radiodurans
dc.typeArticle

Dosyalar