Purification of the Flavoproteins 6-Hydroxy-D- and 6-Hydroxy-L-nicotine Oxidase Using Hydrophobic Affinity Chromatography

Abstract

A systematic study of the homologous series of .omega.-aminoalkyl-agaroses revealed differences in the affinities of [Arthrobacter oxydans] 6-hydroxy-D- and 6-hydroxy-L-nicotine oxidase. In contrast to supports with nonpolar alkyl chains, .omega.-aminoalkyl-agarose showed high affinity towards the L-specific enzyme, while the D-specific oxidase was bound most firmly by .omega.-aminododecyl-agarose. 6-Hydroxy-L-nicotine oxidase could be desorbed by 1.3 M NaCl only in the presence of the substrate L-6-hydroxynicotine. Using the .omega.-aminoalkyl-agarose, a complete separation of the enantiozymes was accomplished and an efficient purification procedure for both oxidases established.