Serum amine oxidase can specifically recognize and oxidize aminohexyl (AH) chains on AH-Sepharose support: single-step affinity immobilization.
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.
Department of Biochemical Sciences 'A. Rossi Fanelli' and CNR Center of Molecular Biology, Rome University 'La Sapienza', 00185-Rome, Italy.
O
O
Befani
M T
MT
Graziani
E
E
Agostinelli
E
E
Grippa
B
B
Mondovì
M A
MA
Mateescu
D006631:Amine Oxidase (Copper-Containing); D000818:Animals; D001596:Benzylamines / Q000378:metabolism; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D004800:Enzymes, Immobilized / Q000302:isolation & purification*; D005650:Fumarates / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000378:metabolism; D007525:Isoelectric Focusing; D008298:Maleates / Q000378:metabolism; D010084:Oxidation-Reduction; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism; D012685:Sepharose / Q000031:analogs & derivatives* / Q000378:metabolism; D013997:Time Factors
D001596:Benzylamines; D004800:Enzymes, Immobilized; D005650:Fumarates; D008298:Maleates; D010455:Peptides; C073341:citraconic acid; C024903:aminohexyl-sepharose; D012685:Sepharose; C030796:benzylamine; D006861:Hydrogen Peroxide; D006631:Amine Oxidase (Copper-Containing); D000587:Oxidoreductases Acting on CH-NH Group Donors