Characterization of H 2O-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 journalArticle

28 Citations (Scopus)

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 °C, respectively, with a K m of 27.0 μM and a k cat/K m of 1.1 × 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.

Original languageEnglish
Pages (from-to)1931-1935
Number of pages5
JournalBioorganic and Medicinal Chemistry Letters
Volume22
Issue number5
DOIs
Publication statusPublished - 2012 Mar 1
Externally publishedYes

Fingerprint

Streptococcus pyogenes
Sugars
NAD
Mutagenesis
Site-Directed Mutagenesis
Escherichia coli
Regeneration
Cats
Temperature
Enzymes
NADH oxidase

Keywords

  • Cofactor regeneration
  • H O-forming NADH oxidase
  • l-Rare sugar
  • Streptococcus pyogenes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmaceutical Science
  • Drug Discovery
  • Clinical Biochemistry
  • Organic Chemistry

Cite this

Characterization of H 2O-forming NADH oxidase from Streptococcus pyogenes and its application in l-rare sugar production. / Gao, Hui; Tiwari, Manish Kumar; Kang, Yun Chan; Lee, Jung Kul.

In: Bioorganic and Medicinal Chemistry Letters, Vol. 22, No. 5, 01.03.2012, p. 1931-1935.

Research output: Contribution to journalArticle

@article{ee6ac3a7a7d24c8983a9ca3ac21c9d5b,
title = "Characterization of H 2O-forming NADH oxidase from Streptococcus pyogenes and its application in l-rare sugar production",
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 °C, respectively, with a K m of 27.0 μM and a k cat/K m of 1.1 × 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.",
keywords = "Cofactor regeneration, H O-forming NADH oxidase, l-Rare sugar, Streptococcus pyogenes",
author = "Hui Gao and Tiwari, {Manish Kumar} and Kang, {Yun Chan} and Lee, {Jung Kul}",
year = "2012",
month = "3",
day = "1",
doi = "10.1016/j.bmcl.2012.01.049",
language = "English",
volume = "22",
pages = "1931--1935",
journal = "Bioorganic and Medicinal Chemistry Letters",
issn = "0960-894X",
publisher = "Elsevier Limited",
number = "5",

}

TY - JOUR

T1 - Characterization of H 2O-forming NADH oxidase from Streptococcus pyogenes and its application in l-rare sugar production

AU - Gao, Hui

AU - Tiwari, Manish Kumar

AU - Kang, Yun Chan

AU - Lee, Jung Kul

PY - 2012/3/1

Y1 - 2012/3/1

N2 - 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 °C, respectively, with a K m of 27.0 μM and a k cat/K m of 1.1 × 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.

AB - 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 °C, respectively, with a K m of 27.0 μM and a k cat/K m of 1.1 × 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.

KW - Cofactor regeneration

KW - H O-forming NADH oxidase

KW - l-Rare sugar

KW - Streptococcus pyogenes

UR - http://www.scopus.com/inward/record.url?scp=84857367018&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84857367018&partnerID=8YFLogxK

U2 - 10.1016/j.bmcl.2012.01.049

DO - 10.1016/j.bmcl.2012.01.049

M3 - Article

VL - 22

SP - 1931

EP - 1935

JO - Bioorganic and Medicinal Chemistry Letters

JF - Bioorganic and Medicinal Chemistry Letters

SN - 0960-894X

IS - 5

ER -