A method for the characterization of aldehyde dehydrogenase with use of alcohol dehydrogenase.
The metabolism of alcohols and glycols is catalyzed sequentially by alcohol and aldehyde dehydrogenases to toxic oxidation products. The study of aldehyde dehydrogenase (ALDH) is complicated by instability and limited availability of aldehyde substrates. We have synthesized aldehydes by use of alcohol dehydrogenase (ADH)-catalyzed oxidation of alcohols; aldehydes produced were immediately used as substrates for ALDH measurements on a centrifugal analyzer. The accuracy of the ADH-ALDH assay was investigated by comparison of the reaction kinetics determined by this method with those obtained using commercially available aliphatic aldehydes as substrates. The mean Michaelis constants determined by the two methods were identical for butyraldehyde and propionaldehyde (3.7 and 4.6 mumol/L, respectively). For glycoaldehyde, values agreed within experimental error: 670 +/- 50 mumol/L and 720 +/- 20 mumol/L using commercial substrate and the proposed method, respectively. This technique allows kinetic studies of the oxidation of a wide variety of toxic aldehydes.
Herold
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DA
Department of Pathology, University of Virginia, Charlottesville 22908.
Keil
K
K
Bruns
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D000426:Alcohol Dehydrogenase; D000438:Alcohols; D000444:Aldehyde Dehydrogenase / Q000032:analysis*; D000447:Aldehydes; D006018:Glycols; D007700:Kinetics; D009243:NAD; D010084:Oxidation-Reduction; D013056:Spectrophotometry, Ultraviolet
D000438:Alcohols; D000447:Aldehydes; D006018:Glycols; D009243:NAD; D000426:Alcohol Dehydrogenase; D000444:Aldehyde Dehydrogenase