Premium
Steroidumwandelnde Enzyme aus Mikroorganismen X. Anreicherung einer 4‐En‐3‐oxosteroid‐5α‐Reduktase aus Mycobacterium smegmatis sowie Abtrennung und Anreicherung des Apoenzyms mit Hilfe der Affinitätschromatographie
Author(s) -
Atrat P.,
Deppmeyer V.,
Groh H.,
Hörhold C.
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.19790190602
Subject(s) - chemistry , mycobacterium smegmatis , enzyme , substrate (aquarium) , urea , elution , chromatography , biochemistry , biology , mycobacterium tuberculosis , medicine , tuberculosis , ecology , pathology
The 4‐en‐3‐oxosteroid‐5α‐reductase from Mycobacterium smegmatis was bound biospecifically on the affinant containing an immobilized testosterone ligand. The enzyme obtained by elution with ethylene glycol and urea in a 32 fold purity has a S. A. of 8.73 × 10 −3 μM androstenedione min −1 mg −1 . The coenzyme (FAD) could be separated from the immobilized enzyme substrate complex on the affinity matrix, in the presence of (NH 4 ) 2 SO 4 at pH 3.0. After elution of the apoenzyme 97% of the initial enzyme activity was obtained by incubation with FAD. The reactivated enzyme results in a 40 fold enrichment.