Purification of the flavoproteins 6-hydroxy-D- and 6-hydroxy-L-nicotine oxidase using hydrophobic affinity chromatography.
A systematic study of the homologous series of omega-aminoalkyl-agaroses revealed differences in the affinities of 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 1.3M 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.
D001173:Arthrobacter / Q000201:enzymology; D002846:Chromatography, Affinity / Q000379:methods; D008970:Molecular Weight; D010088:Oxidoreductases / Q000302:isolation & purification*; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000302:isolation & purification*
D010088:Oxidoreductases; D000587:Oxidoreductases Acting on CH-NH Group Donors; C037584:6-hydroxy-L-nicotine oxidase; C021413:6-hydroxy-D-nicotine oxidase