Steroidumwandelnde enzyme aus mikroorganismen—VII. Untersuchungen zur natur der biospezifischen bindung an der affinitätsmatrix—affinitätschromatographie einer 4-en-3-oxo-steroid: (Akzeptor)-1-en-oxidoreduktase aus Nocardia opaca

Abstract

For affinity chromatography of the 4-en-3-oxosteroid: (acceptor)-1-en-oxidoreductase (EC 1.3.99.4.) from Nocardia opaca were applied two affinants with different spacers between the immobilized testosterone ligand and matrix. Affinant 1 contains hexamethylenediamine and affinant 2 adipinic acid dihydrazide as the spacer. On both affinants the enzyme was bound biospecifically. These affinants show a different elution behaviour. From the effect of potassium chloride and such organic solvents as glycerol, ethyleneglycol and dimethylformamide the biospecific binding on the immobilized testosterone ligand was concluded to be hydrophobic. This interchange can be influenced by the nature of spacer and spacermatrix-binding in the case of a given ligand density and a constant ligand-matrix distance.