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

doi:10.1016/j.bmcl.2012.01.049

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 journal › Article › peer-review

42 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 language	English
Pages (from-to)	1931-1935
Number of pages	5
Journal	Bioorganic and Medicinal Chemistry Letters
Volume	22
Issue number	5
DOIs	https://doi.org/10.1016/j.bmcl.2012.01.049
Publication status	Published - 2012 Mar 1
Externally published	Yes

Bibliographical note

Funding Information:
This work was supported by the 21C Frontier Microbial Genomics and Applications Center Program, Ministry of Education, Science & Technology, Republic of Korea . This research was also supported by the Converging Research Center Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology ( 2011-50210 ).

Keywords

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

ASJC Scopus subject areas

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

Access to Document

10.1016/j.bmcl.2012.01.049

Cite this

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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.",

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

note = "Funding Information: This work was supported by the 21C Frontier Microbial Genomics and Applications Center Program, Ministry of Education, Science & Technology, Republic of Korea . This research was also supported by the Converging Research Center Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology ( 2011-50210 ). ",

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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

N1 - Funding Information: This work was supported by the 21C Frontier Microbial Genomics and Applications Center Program, Ministry of Education, Science & Technology, Republic of Korea . This research was also supported by the Converging Research Center Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology ( 2011-50210 ).

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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 - Streptococcus pyogenes

KW - l-Rare sugar

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