The carboxylic acid reduction pathway in Nocardia. Purification and characterization of the aldehyde reductase

Whole cultures of Nocardia sp. NRRL 5646 reduce carboxylic acids, first to aldehydes, then to alcohols and subsequently to the corresponding acetyl esters. This work describes an NADPH-dependent reductase responsible for catalyzing the reduction of aldehyde intermediates, which was purified 3240-fold by a combination of Mono-Q, hydroxyapatite, and ADP-agarose chromatographies. By sodium dodecyl sulfate–polyacrylamide gel electrophoresis, the purified enzyme ran as a single band of 47 kDa. A native molecular mass estimated at 101 kDa indicated that the enzyme was a homodimer in the native, active state. Edman degradation indicated a unique N-terminal sequence as NH₂-X-X-Ala-Ala-Ala-Tyr-Ala-Val-Pro-Ala-Pro-Asp-Gly-Cys-Phe-Glu-Lys-Val-Thr-Ile-Glu-Arg-Arg-Glu-Leu-Gly. The enzyme catalyzed reductions of many aryl- and alkyl-aldehyde substrates. Reactions were most favorable in the direction of aldehyde reduction to alcohols. Journal of Industrial Microbiology & Biotechnology (2000) 25, 328–332.