NADH oxidase (Nox) catalyzes the conversion of NADH to NAD<SUP>+</SUP>. A previously uncharacterized Nox gene (LrNox) was cloned from Lactobacillus rhamnosus and overexpressed in Escherichia coli BL21(DE3). Sequence analysis revea...
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https://www.riss.kr/link?id=A107626513
Zhang, Y.W. ; Tiwari, M.K. ; Gao, H. ; Dhiman, S.S. ; Jeya, M. ; Lee, J.K.
2012
-
SCOPUS,SCIE
학술저널
255-262(8쪽)
0
상세조회0
다운로드다국어 초록 (Multilingual Abstract)
NADH oxidase (Nox) catalyzes the conversion of NADH to NAD<SUP>+</SUP>. A previously uncharacterized Nox gene (LrNox) was cloned from Lactobacillus rhamnosus and overexpressed in Escherichia coli BL21(DE3). Sequence analysis revea...
NADH oxidase (Nox) catalyzes the conversion of NADH to NAD<SUP>+</SUP>. A previously uncharacterized Nox gene (LrNox) was cloned from Lactobacillus rhamnosus and overexpressed in Escherichia coli BL21(DE3). Sequence analysis revealed an open reading frame of 1359bp, capable of encoding a polypeptide of 453 amino acid residues. The molecular mass of the purified LrNox enzyme was estimated to be ∼50kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and 100kDa by gel filtration chromatography, suggesting that the enzyme is a homodimer. The enzyme had optimal activity at pH 5.6 and temperature 65<SUP>o</SUP>C, and k<SUB>cat</SUB>/K<SUB>m</SUB> of 3.77x10<SUP>7</SUP>s<SUP>-1</SUP>M<SUP>-1</SUP>, the highest ever reported. Heat inactivation studies revealed that LrNox had high thermostability, with a half-life of 120min at 80<SUP>o</SUP>C. Molecular dynamics simulation studies shed light on the factors contributing to the high activity of LrNox. Although the properties of Nox from several microorganisms have been reported, this is the first report on the characterization of a recombinant H<SUB>2</SUB>O-forming Nox with high activity and thermostability. The characteristics of the LrNox enzyme could prove to be of interest in industrial applications such as NAD<SUP>+</SUP> regeneration.