Characterization of H2O-forming NADH oxidase from Streptococcus pyogenes and its application in L-rare sugar production

Hui Gao, Manish Kumar Tiwari, Yun Chan Kang, Jung-Kul Lee

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

A nicotinamide adenine dinucleotide (NADH) oxidase from Streptococcus pyogenes MGAS10394 (SpNox) was cloned and overexpressed in Escherichia coli BL21 (DE3). The purified SpNox enzyme had optimal pH and temperature of 7.0 and 55 degrees C, respectively, with a K-m of 27.0 mu M and a k(cat)/K-m of 1.1 x 10(7) s (1) M (1). SpNox showed the highest activity among all known NADH oxidases, and site-directed mutagenesis and docking analysis shed light on the molecular basis of its unusually high activity. The characteristics of SpNox may prove to be useful for NAD(+) regeneration in the production of L-rare sugar. (c) 2012 Elsevier Ltd. All rights reserved.
Original languageEnglish
JournalBioorganic & Medicinal Chemistry Letters
Volume22
Issue number5
Pages (from-to)1931-1935
ISSN0960-894X
DOIs
Publication statusPublished - 2012
Externally publishedYes

Keywords

  • Cloning, Molecular
  • Crystallography, X-Ray
  • Escherichia coli
  • Models, Molecular
  • Multienzyme Complexes
  • Mutagenesis, Site-Directed
  • NADH, NADPH Oxidoreductases
  • Streptococcus pyogenes
  • Xylulose
  • 5962-29-8 Xylulose
  • EC 1.6.- NADH oxidase
  • EC 1.6.- NADH, NADPH Oxidoreductases
  • reduced nicotinamide adenine dinucleotide dehydrogenase
  • sugar
  • multienzyme complex
  • oxidoreductase
  • xylulose
  • article
  • catalysis
  • enzyme metabolism
  • enzyme structure
  • molecular docking
  • nonhuman
  • pH
  • site directed mutagenesis
  • temperature
  • chemical structure
  • chemistry
  • enzymology
  • genetics
  • metabolism
  • molecular cloning
  • X ray crystallography
  • Cofactor regeneration
  • H 2O-forming NADH oxidase
  • l-Rare sugar
  • enzyme activity
  • Eubacteria Bacteria Microorganisms (Bacteria, Eubacteria, Microorganisms) - Gram-Positive Cocci [07700] Streptococcus pyogenes species strain-MGAS10394
  • Facultatively Anaerobic Gram-Negative Rods Eubacteria Bacteria Microorganisms (Bacteria, Eubacteria, Microorganisms) - Enterobacteriaceae [06702] Escherichia coli species strain-BL21
  • L-rare sugar
  • NAD 53-84-9
  • nicotinamide adenine dinucleotide oxidase NADH oxidase expression EC 1.6.99.3
  • water 7732-18-5
  • 10060, Biochemistry studies - General
  • 10062, Biochemistry studies - Nucleic acids, purines and pyrimidines
  • 31000, Physiology and biochemistry of bacteria
  • 39008, Food microbiology - General and miscellaneous
  • cloning laboratory techniques, genetic techniques
  • docking analysis laboratory techniques
  • site-directed mutagenesis laboratory techniques, genetic techniques
  • Biochemistry and Molecular Biophysics
  • Bioprocess Engineering
  • CHEMISTRY,
  • FLAVOPROTEIN DISULFIDE REDUCTASES
  • GLUTATHIONE-REDUCTASE
  • BACILLUS-PALLIDUS
  • PURIFICATION
  • CONVERSION
  • CATALYSIS
  • COFACTOR
  • FAECALIS
  • CLONING
  • ENZYME
  • H2O-forming NADH oxidase
  • L-Rare sugar

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