Molecular cloning and characterization of NAD(+) dependent isocitrate dehydrogenase enzyme from Shewanella putrefaciens

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Fahri A., Metin D., Ahmet O., Sahabettin S.

RESEARCH JOURNAL OF BIOTECHNOLOGY, cilt.16, sa.10, ss.78-86, 2021 (ESCI, Scopus) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 16 Sayı: 10
  • Basım Tarihi: 2021
  • Dergi Adı: RESEARCH JOURNAL OF BIOTECHNOLOGY
  • Derginin Tarandığı İndeksler: Emerging Sources Citation Index (ESCI), Scopus
  • Sayfa Sayıları: ss.78-86
  • Anahtar Kelimeler: Cloning, enzyme activity, isocitrate dehydrogenase, recombinant enzyme, Shewanella putrefaciens, BIOCHEMICAL-CHARACTERIZATION, COFACTOR SPECIFICITY, PURIFICATION, BACTEREMIA, MUTATIONS, REVEALS, PATIENT, ALGAE
  • Açık Arşiv Koleksiyonu: AVESİS Açık Erişim Koleksiyonu
  • İstanbul Üniversitesi-Cerrahpaşa Adresli: Evet

Özet

Isocitrate dehydrogenase (IDH) is a fundamental enzyme for carbon metabolism in the Krebs cycle. This enzyme is required for oxidation-reduction reactions in both eukaryotic and prokaryotic cells and plays a critical role in their growth and pathogenesis. In this study, we cloned the gene encoding NAD(+) dependent isocitrate dehydrogenase from Shewanella putrefaciens. The expression of recombinant protein was induced with 0.5 mM of IPTG. His-tagged IDH overexpressed in E. coli was purified and characterized. The expressed IDH enzyme was purified in an active soluble form. The molecular weight of the enzyme was confirmed with Western blotting. High sequence homology was observed with IDH sequences of other Shewanella strains and remarkable sequence homology was found with other bacteria reported in the database.