Properties of delta5-3beta-hydroxysteroid oxidoreductase isolated from Streptomyces griseocarneus.

G. Kerényi, A. Szentirmai, M. Natonek

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Abstract

Delta5-3beta-hydroxysteroid oxidoreductase was extracted in magnesium-containing Tris buffer from sonicated Streptomyces griseocarneus cells. The enzyme was partially purified (150 X) by ion exchange chromatography and gel filtration following (NH4)2SO4 fractionation. Upon gel filtration on Sephadex G-75 to G-200, the greatest part of the activity gave a peak in the fractionation range. The enzyme obtained from the gel yielded small enzyme molecules on repeated chromatography. A molecular weight of 32 to 36 000 was calculated for the activity appearing in the fractionation range of Sephadex G-75 to G-200. The enzyme is highly specific for the irreversible oxidation of the 3beta-hydroxyl group in steroids with a trans-anellated A : B ring system with either C5 or C6 double bond. Delta5-3-ketosteroids are converted into delta5-3-ketosteroids at a high rate, but the isomerase activity cannot be separated from the oxidoreductase activity either by chromatography or by selective heat inactivation. NAD, NADP, FMN or FAD did not influence the activity, but the enzyme is inactive in the absence of molecular oxygen.

Original languageEnglish
Pages (from-to)487-496
Number of pages10
JournalActa Microbiologica Academiae Scientiarum Hungaricae
Volume22
Issue number4
Publication statusPublished - 1975

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Hydroxysteroids
Streptomyces
Oxidoreductases
Ketosteroids
Enzymes
Gel Chromatography
Chromatography
Flavin Mononucleotide
Isomerases
Tromethamine
Flavin-Adenine Dinucleotide
Ion Exchange Chromatography
NADP
Hydroxyl Radical
NAD
Magnesium
Hot Temperature
Molecular Weight
Gels
Steroids

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Properties of delta5-3beta-hydroxysteroid oxidoreductase isolated from Streptomyces griseocarneus. / Kerényi, G.; Szentirmai, A.; Natonek, M.

In: Acta Microbiologica Academiae Scientiarum Hungaricae, Vol. 22, No. 4, 1975, p. 487-496.

Research output: Contribution to journalArticle

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