[Steroid-transforming enzymes from microorganisms. X. Enrichment of a 4-en-3-oxosteroid-5 alpha-reductase from Mycobacterium smegmatis as well as separation and enrichment of the apoenzyme by means of affinity chromatography].
The 4-en-3-oxosteroid-5 alpha-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 X 10(-3) microM 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 (NH4)2SO4 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.
D013741:3-Oxo-5-alpha-Steroid 4-Dehydrogenase / Q000302:isolation & purification* / Q000378:metabolism; D001051:Apoenzymes / Q000302:isolation & purification* / Q000378:metabolism; D001059:Apoproteins / Q000302:isolation & purification*; D002474:Cell-Free System; D002846:Chromatography, Affinity; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism; D009161:Mycobacterium / Q000201:enzymology*; D010088:Oxidoreductases / Q000302:isolation & purification*
D001051:Apoenzymes; D001059:Apoproteins; D005182:Flavin-Adenine Dinucleotide; D010088:Oxidoreductases; D013741:3-Oxo-5-alpha-Steroid 4-Dehydrogenase
Steroidumwandelnde Enzyme aus Mikroorganismen. X. Anreicherung einer 4-En-3-oxosteroid-5 alpha-Reduktase aus Mycobacterium smegmatis sowie Abtrennung und Anreicherung des Apoenzyms mit Hilfe der Affinitätschromatographie.