Hyperthermophilic flavin reductase from Sulfolobus solfataricus P2: Production and biochemical characterization

Gun G., Imamoglu R., Tatli O., Yurum Y., BAYKAL A. T., Dinler-Doganay G.

BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY, cilt.66, sa.6, ss.915-923, 2019 (SCI-Expanded) identifier identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 66 Sayı: 6
  • Basım Tarihi: 2019
  • Doi Numarası: 10.1002/bab.1801
  • Dergi Adı: BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.915-923
  • Anahtar Kelimeler: archaea, biodesulfurization, flavin reductase activity, hyperthermophile, oxidoreductase, NAD(P)H-FLAVIN OXIDOREDUCTASE, DESULFURIZING BACTERIUM, ESCHERICHIA-COLI, DIBENZOTHIOPHENE, PURIFICATION, BIODESULFURIZATION, SYSTEM, MECHANISMS
  • Acıbadem Mehmet Ali Aydınlar Üniversitesi Adresli: Evet

Özet

Nicotinamide adenine dinucleotide phosphate (NAD(P)H)-flavin oxidoreductases (flavin reductases) catalyze the reduction of flavin by NAD(P)H and provide the reduced form of flavin mononucleotide (FMN) to flavin-dependent monooxygenases. Based on bioinformatics analysis, we identified a putative flavin reductase gene, sso2055, in the genome of hyperthermophilic archaeon Sulfolobus solfataricus P2, and further cloned this target sequence into an expression vector. The cloned flavin reductase (EC. 1.5.1.30) was purified to homogeneity and characterized further. The purified enzyme exists as a monomer of 17.8 kDa, free of chromogenic cofactors. Homology modeling revealed this enzyme as a TIM barrel, which is also supported by circular dichroism measurements revealing a beta-sheet rich content. The optimal pH for SSO2055 activity was pH 6.5 in phosphate buffer and the highest activity observed was at 120 degrees C within the measurable temperature. We showed that this enzyme can use FMN and flavin adenine dinucleotide (FAD) as a substrate to generate their reduced forms. The purified enzyme is predicted to be a potential flavin reductase of flavin-dependent monooxygenases that could be involved in the biodesulfurization process of S. solfataricus P2.