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

archaea biodesulfurization flavin reductase activity hyperthermophile oxidoreductase

Journal

Biotechnology and applied biochemistry
ISSN: 1470-8744
Titre abrégé: Biotechnol Appl Biochem
Pays: United States
ID NLM: 8609465

Informations de publication

Date de publication:
Nov 2019
Historique:
received: 20 05 2019
accepted: 03 08 2019
pubmed: 10 8 2019
medline: 25 3 2020
entrez: 10 8 2019
Statut: ppublish

Résumé

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 °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.

Identifiants

pubmed: 31396993
doi: 10.1002/bab.1801
doi:

Substances chimiques

Oxidoreductases EC 1.-

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

915-923

Subventions

Organisme : Türkiye Bilimsel ve Teknolojik Araştirma Kurumu
ID : 110M001

Informations de copyright

© 2019 International Union of Biochemistry and Molecular Biology, Inc.

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Auteurs

Gokhan Gun (G)

Department of Molecular Biology & Genetics, Istanbul Technical University, Maslak, Istanbul, Turkey.

Rahmi Imamoglu (R)

Department of Molecular Biology & Genetics, Istanbul Technical University, Maslak, Istanbul, Turkey.

Ozge Tatli (O)

Department of Molecular Biology & Genetics, Istanbul Technical University, Maslak, Istanbul, Turkey.
Department of Molecular Biology & Genetics, Istanbul Medeniyet University, Istanbul, Turkey.

Yuda Yurum (Y)

Department of Molecular Biology & Genetics, Sabanci University, Istanbul, Turkey.

Ahmet Tarik Baykal (A)

Department of Medical Biochemistry, School of Medicine, Acibadem University, Istanbul, Turkey.

Gizem Dinler-Doganay (G)

Department of Molecular Biology & Genetics, Istanbul Technical University, Maslak, Istanbul, Turkey.

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