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Biotechnology and Applied Biochemistry (1998) 28, (99–104) (Printed in Great Britain)

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Serum amine oxidase can specifically recognize and oxidize aminohexyl (AH) chains on AH-Sepharose support: single-step affinity immobilization¹

Olivia Befani*, Maria Teresa Graziani*², Enzo Agostinelli*, Eleonora Grippa*, Bruno Mondovì* and Mircea-Alexandru Mateescu†³

*Department of Biochemical Sciences 'A. Rossi Fanelli' and CNR Center of Molecular Biology, Rome University 'La Sapienza', 00185 – Rome, Italy, and †Department of Chemistry–Biochemistry, Université du Québec à Montréal, CP 8888, Succ. A, Montréal, Québec, Canada H3C 3P8

¹ This paper is dedicated to the memory of Maria Teresa Graziani.

² Deceased.

³ To whom correspondence should be addressed.

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.

Received 28 May 1998; accepted 25 June 1998

Portland Press Ltd © 1998