Coupled-enzyme determination of catalase activity in erythrocytes

Autor:	Michael Pepoy, F Van Lente
Rok vydání:	1990
Předmět:	chemistry.chemical_classification Chromatography Ethanol biology Biochemistry (medical) Clinical Biochemistry Acetaldehyde Substrate (chemistry) Enzyme assay chemistry.chemical_compound Enzyme chemistry Biochemistry Catalase biology.protein Glucose oxidase Hydrogen peroxide
Zdroj:	Clinical Chemistry. 36:1339-1343
ISSN:	1530-8561 0009-9147
Popis:	This coupled-enzyme method for determining the activity of catalase (EC 1.11.1.6) in erythrocyte lysates is based on measuring the absorbance at 340 nm of NADH produced from the peroxidic reaction between ethanol, hydrogen peroxide, and catalase. Hydrogen peroxide is produced as a substrate in situ from the oxidation of glucose catalyzed by glucose oxidase (EC 1.1.3.4). Catalase oxidizes ethanol to acetaldehyde in the presence of hydrogen peroxide. Acetaldehyde is then oxidized by aldehyde dehydrogenase (EC 1.2.1.5) to produce acetate with concomitant conversion of NAD+ to NADH. The reaction did not follow strict zero-order kinetics; enzyme activity was quantified by using initial rates and standards prepared from purified catalase. The method demonstrated within-run and between-run CVs of 1.0% to 2.9% and 2.4% to 3.3%, respectively. This semiautomated method correlated well (r = 0.92) with the more tedious manual method involving measurement at 240 nm.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::5dccbfedbb120cd4b422e40bf731aee7 https://doi.org/10.1093/clinchem/36.7.1339 Zobrazit plný text záznamu