Befani O, Graziani M T, Agostinelli E, Grippa E, Mondovì B, Mateescu M A
Department of Biochemical Sciences 'A. Rossi Fanelli' and CNR Center of Molecular Biology, Rome University 'La Sapienza', 00185-Rome, Italy.
Biotechnol Appl Biochem. 1998 Oct;28 ( Pt 2):99-104.
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.
牛血清胺氧化酶（SAO）在氨基己基（AH）-琼脂糖凝胶上的制备性亲和层析常常伴随着SAO在载体上意外的不可逆保留。这种不使用偶联试剂的特殊酶固定化被认为是由于SAO具有以下能力：（i）将载体的烷基胺基团识别为大分子化的底物；（ii）催化它们氧化为相应的醛，同时释放出NH₃和H₂O₂；以及（iii）通过新生醛基与SAO游离胺基之间的偶联固定在活化载体上。这种具有载体自活化作用的亲和固定化程序较为温和，通过简单孵育24小时，就能实现酶的固定化，且保留了游离SAO原始比活性的80%。固定在AH-琼脂糖凝胶微柱上的SAO，被视为连续流动系统反应器，能够在数周内催化苄胺氧化。
