sepharose and benzylamine

Preparative affinity chromatography of bovine serum amine oxidase (SAO) on aminohexyl (AH)-Sepharose was often associated with an unexpected irreversible SAO retention on the support. This particular enzyme immobilization, occurring without coupling reagents, was supposed to be due to a SAO ability to: (i) recognize alkylamine groups of the support as macro-molecularized substrate; (ii) catalyse their oxidation to the corresponding aldehydes, with release of NH3 and H2O2; and (iii) be immobilized on the activated support by a coupling between the nascent aldehyde groups and SAO free amine groups. This affinity immobilization procedure, with the self-activation of the support, being mild, allows by simple incubation for 24 h, the enzyme immobilization with the retention of 80% from original specific activity of free SAO. Immobilized SAO on AH-Sepharose microcolumns, viewed as a continuous flow-system reactor, was able to catalyse benzylamine oxidation for several weeks.

Topics: Amine Oxidase (Copper-Containing); Animals; Benzylamines; Cattle; Chromatography, Affinity; Enzymes, Immobilized; Fumarates; Hydrogen Peroxide; Isoelectric Focusing; Maleates; Oxidation-Reduction; Oxidoreductases Acting on CH-NH Group Donors; Peptides; Sepharose; Time Factors