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Steroidumwandelnde Enzyme aus Mikroorganismen IX. Affinitätschromatographische Präparation und Untersuchung des Apoenzyms einer 4‐En‐3‐oxosteroid: (Akzeptor)‐1‐en‐oxidoreduktase aus Nocardia opaca
Author(s) -
Atrat P.,
Deppmeyer V.,
Hórhold C.,
Móller L.
Publication year - 1979
Publication title -
zeitschrift für allgemeine mikrobiologie
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0044-2208
DOI - 10.1002/jobm.19790190502
Subject(s) - chemistry , enzyme , ligand (biochemistry) , elution , stereochemistry , chromatography , biochemistry , receptor
From the flavoenzyme, 4‐en‐3‐oxosteroid: (acceptor)‐1‐en‐oxidoreductase of Nocardia opaca , prosthetic group and apoenzyme were separated quantitatively by means of affinity chromatography in the presence of 2 M (NH 4 ) 2 SO 4 at pH 3.0. Subsequently the apoenzyme was eluted from affinity matrix by 0.01 M phosphate buffer, pH 8.0, whereas under these conditions the intact enzyme could not be eluted. The whole enzyme activity applied could be restored by incubation of the eluted apoenzyme with FAD. The binding strength of the apoenzyme to the immobilized steroid ligand is highly decreased in comparison to the native enzyme and can be interpreted by the action of rest hydrophobicity. That indicates the essential character of FAD for both ligand binding and transformation.