Premium
Serum amine oxidase can specifically recognize and oxidize aminohexyl (AH) chains on AH‐Sepharose support: single‐step affinity immobilization 1
Author(s) -
Befani Olivia,
Graziani Maria Teresa,
Agostinelli Enzo,
Grippa Eleonora,
Mondovì Bruno,
Mateescu MirceaAlexandru
Publication year - 1998
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1998.tb00518.x
Subject(s) - sepharose , chemistry , amine oxidase , amine gas treating , affinity chromatography , enzyme , biochemistry , chromatography , stereochemistry , combinatorial chemistry , organic chemistry
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 NH 3 and H 2 O 2 ; 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.