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

doi:10.1016/j.bmcl.2012.01.049

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 journal › Journal article › Research › peer-review

46 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 27.0μM 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 language	English
Journal	Bioorganic & Medicinal Chemistry Letters
Volume	22
Issue number	5
Pages (from-to)	1931–1935
Number of pages	5
ISSN	0960-894X
DOIs	https://doi.org/10.1016/j.bmcl.2012.01.049
Publication status	Published - 2012
Externally published	Yes

Keywords

Cloning, Molecular
Crystallography, X-Ray
Escherichia coli
Models, Molecular
Multienzyme Complexes
Mutagenesis, Site-Directed
NADH, NADPH Oxidoreductases
Streptococcus pyogenes
Xylulose

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Cite this

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title = "Characterization of H2O-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 27.0μM 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 = "Cloning, Molecular, Crystallography, X-Ray, Escherichia coli, Models, Molecular, Multienzyme Complexes, Mutagenesis, Site-Directed, NADH, NADPH Oxidoreductases, Streptococcus pyogenes, Xylulose",

author = "Hui Gao and Tiwari, {Manish Kumar} and Kang, {Yun Chan} and Jung-Kul Lee",

year = "2012",

doi = "10.1016/j.bmcl.2012.01.049",

language = "English",

volume = "22",

pages = "1931–1935",

journal = "Bioorganic & Medicinal Chemistry Letters",

issn = "0960-894X",

publisher = "Pergamon Press",

number = "5",

}

TY - JOUR

T1 - Characterization of H2O-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

Y1 - 2012

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 27.0μM 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 27.0μM 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 - Cloning, Molecular

KW - Crystallography, X-Ray

KW - Escherichia coli

KW - Models, Molecular

KW - Multienzyme Complexes

KW - Mutagenesis, Site-Directed

KW - NADH, NADPH Oxidoreductases

KW - Streptococcus pyogenes

KW - Xylulose

U2 - 10.1016/j.bmcl.2012.01.049

DO - 10.1016/j.bmcl.2012.01.049

M3 - Journal article

C2 - 22326164

SN - 0960-894X

VL - 22

SP - 1931

EP - 1935

JO - Bioorganic & Medicinal Chemistry Letters

JF - Bioorganic & Medicinal Chemistry Letters

IS - 5

ER -