Taylor Russell H, Fournier Sylvie M, Simons Brigitte L, Kaplan Harvey, Hefford Mary Alice
Department of Biochemistry, University of Ottawa, Canada K1H 8M5.
J Biotechnol. 2005 Aug 22;118(3):265-9. doi: 10.1016/j.jbiotec.2005.05.007.
Lyophilized alkaline phosphatase (ALPase) was immobilized on aminated glass surfaces using the in vacuo cross-linking process [Simons, B.L., King, M.C., Cyr, T., Hefford, M.A., Kaplan, H., 2002. Zero-length cross-linking of lyophilized proteins. Protein Sci. 11, 1558-1564]. In this procedure, amide bonds were formed between carboxyl groups on the protein and amino groups on the glass surface. After the non-covalently attached enzyme was removed the immobilized ALPase not only retained its activity but could also be used, washed and reused at least six times without significant loss of activity. An average of 1.4+/-0.6 mg of reusable ALPase per gram of glass fibre was immobilized based on the activity of the soluble equivalent.
采用真空交联法将冻干碱性磷酸酶（ALPase）固定在胺化玻璃表面[西蒙斯，B.L.，金，M.C.， Cyr，T.，赫福德，M.A.，卡普兰，H.，2002年。冻干蛋白质的零长度交联。蛋白质科学。11，1558 - 1564]。在此过程中，蛋白质上的羧基与玻璃表面的氨基之间形成酰胺键。去除非共价结合的酶后，固定化的ALPase不仅保留了其活性，而且至少可使用、洗涤和重复使用六次，活性无显著损失。基于可溶性等效物的活性，每克玻璃纤维平均固定化1.4±0.6毫克可重复使用的ALPase。
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