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Journals / Hacettepe Journal of Biology and Chemistry / 2021 / Cilt: 49 - Sayı: 3

Cloning, Over-Expression, and Purification of β–Carbonic Anhydrase from an Extremophilic Bacterium: Deinococcus radiodurans

Ayşe Hümeyra TAŞKIN KAFA
Arzu CİHAN
Mehmet KUZUCU
Murat ÇANKAYA
Pages
219–231
Publication Date
2021-05-17
DOI
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AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

The CO2 hydration assay demonstrated that the specific activity of purified recombinant enzymes (DrCA) was significantly high.

📌 3 Key Takeaways

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Objective & Hypothesis:Öz In 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.
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Key Findings & Data:In addition, the effect of increased gamma irradiation doses on pH-related DrCA enzyme activity was determined.
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Conclusion & Impact:The CO2 hydration assay demonstrated that the specific activity of purified recombinant enzymes (DrCA) was significantly high.
🔬Design:Research Article
📍Scope:Comprehensive Sample
⚙️Variables:Anahtar Kelimeler Deinococcus radiodurans · carbonic anhydrase · ionizing radiation

Özet

Öz In 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 CO2 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 Zn2+, which resulted in 5-fold increases of CO2 hydration activity. Also sulfonamide showed inhibitory effect on the pure enzyme. The apparent Km and Vmax for CO2 as substrate were 8.4 mM and 637 WAU/mg for DrCA respectively. The CO2 hydration assay demonstrated that the specific activity of purified recombinant enzymes (DrCA) was significantly high.

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Cite This Article

KAFA, A. H. T., CİHAN, A., KUZUCU, M., ÇANKAYA, M. (2021). Cloning, Over-Expression, and Purification of β–Carbonic Anhydrase from an Extremophilic Bacterium: Deinococcus radiodurans. Hacettepe Journal of Biology and Chemistry, Cilt: 49 - Sayı: 3, 219–231. https://ajindex.com/articles/255632

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