Purification and Properties of a 3α-Hydroxysteroid Dehydrogenase from Pseudomonas testosteroni

Jean Boyer, Denis N. Baron, Paul Talalay

Research output: Contribution to journalArticlepeer-review

36 Scopus citations


A procedure is described for the purification of the steroid-induced 3α-hydroxysteroid dehydrogenase of Pseudomonas testosteroni. The purified preparations catalyze the nicotinamide-adenine dinucleotide dependent oxidation of about 300,umoles of androsterone/min per mg of protein at pH 9.0 and 25°. The purification depends upon stabilization of the enzyme in glycerol-water mixtures; removal of nucleic acids with protamine sulfate; ammonium sulfate and acetone fractionations; chromatography on carboxymethyl-cellulose; and gel filtration on Sephadex G-100. The enzyme reacts also with the thionicotinamide, the 3-pyridinealdehyde, and the acetylpyridine analogs of nicotinamide-adenine dinucleotide. The effect of pH and temperature on the reaction have been measured. The application of the enzyme to the sensitive enzymatic assay of androsterone is illustrated.

Original languageEnglish (US)
Pages (from-to)1825-1833
Number of pages9
Issue number9
StatePublished - Sep 1 1965
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry


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