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Titolo:
Enzymatic assay of dehydrogenase substrate based on the detection of superoxide anion
Autore:
Sarker, AK; Ukeda, H; Kawana, D; Sawamura, M;
Indirizzi:
Kochi Univ, Fac Agr, Dept Bioresource Sci, Nankoku, Kochi 7838502, Japan Kochi Univ Nankoku Kochi Japan 7838502 Sci, Nankoku, Kochi 7838502, Japan
Titolo Testata:
FOOD RESEARCH INTERNATIONAL
fascicolo: 5, volume: 34, anno: 2001,
pagine: 393 - 399
SICI:
0963-9969(2001)34:5<393:EAODSB>2.0.ZU;2-W
Fonte:
ISI
Lingua:
ENG
Soggetto:
FLOW-INJECTION ANALYSIS; WATER-SOLUBLE TETRAZOLIUM; XANTHINE-XANTHINE OXIDASE; NADH-OXIDASE; CARBON ELECTRODES; ELECTROCATALYTIC OXIDATION; SPECTROPHOTOMETRIC ASSAY; LIPOAMIDE DEHYDROGENASE; ALCOHOL-DEHYDROGENASE; REDUCTION;
Keywords:
superoxide anion; tetrazolium salt; NADH oxidase; diaphorase; ethanol determination;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Citazioni:
35
Recensione:
Indirizzi per estratti:
Indirizzo: Ukeda, H Kochi Univ, Fac Agr, Dept Bioresource Sci, Monobe B-200, Nankoku,Kochi 7838502, Japan Kochi Univ Monobe B-200 Nankoku Kochi Japan 7838502 838502, Japan
Citazione:
A.K. Sarker et al., "Enzymatic assay of dehydrogenase substrate based on the detection of superoxide anion", FOOD RES IN, 34(5), 2001, pp. 393-399

Abstract

The oxidation of NADH using NADH oxidase from Bacillus licheniformis or diaphorase from Clostridium kluyveri, was found to produce superoxide anion (O-2(-)). The generated O-2(-) reduced water-soluble tetrazolium salt WST-1 to WST-1 formazan. The formation of WST-1 formazan was monitored as a change in the absorbance at 438 nm. The formation of WST-1 formazan was found tobe proportional to the NADH concentration. As a result, a linear curve wasobtained within the range of 0.5 muM-50 muM NADH concentration. The concentration of NADH was determined by chemiluminescence using lucigenin specific for O-2(-) instead of WST-1. Application of NADH oxidase from Bacillus licheniformis was found to be 400 times stronger chemiluminescence than that of diaphorase with a lower detection limit of 15 nM. The present method wasapplied to determine ethanol combined with yeast alcohol dehydrogenase. The ethanol concentrations of various kinds of alcoholic beverages obtained by the present method were compared with those obtained using F-kit method. A good linear correlation was observed between them (r=0.9997) and the slope (1.005) was very close to unity, suggesting that the present method couldbe applied for the determination of ethanol in practical samples. (C) 2001Elsevier Science Ltd. All rights reserved.

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Documento generato il 29/09/20 alle ore 00:23:36