Abstract
Publisher Summary Aldehyde reductase from the pig kidney is a monomeric oxidoreductase of broad substrate specificity. The enzyme catalyzes the reduction of a wide range of aliphatic and aromatic aldehydes and is similar to another monomeric nicotinamide adenine dinucleotide phosphate dehydrogenase (NADPH)-linked oxidoreductase, aldose reductase. Aldehyde reductase is assayed spectrophotometrically by determining the decrease in the absorbance of NADPH at 340 nm. The reaction is carried out in quartz cuvettes in a double-beam recording spectrophotometer at 25°C. Reaction mixtures contain a final concentration of 100 m M sodium phosphate buffer, pH 7.0, 160 μ M NADPH, 5 m M pyridine-3-aldehyde, and 0.1 ml of enzyme solution in a total volume of 3.0 ml. Reactions are initiated by the addition of enzyme. Blank cuvettes contain all reagents at the same concentration, but with pyridine-3- aldehyde omitted. The steps involved in the purification of the enzyme are also discussed in the chapter. The procedure used is that of Davidson and Flynn with minor modifications. All procedures are carried out at 0–4°C. Aldehyde reductase uses NADPH exclusively as a coenzyme.
Published Version
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