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用于酵母转化酶吸附且可重复使用的伴刀豆球蛋白A固定化亲和吸附剂。

Concanavalin a immobilized affinity adsorbents for reversible use in yeast invertase adsorption.

作者信息

Yavuz Handan, Akgöl Sinan, Arica Yakup, Denizli Adil

机构信息

Department of Chemistry, Hacettepe University, Ankara, Turkey.

出版信息

Macromol Biosci. 2004 Jul 14;4(7):674-9. doi: 10.1002/mabi.200400028.

Abstract

Concanavalin A (Con A) immobilized poly(2-hydroxyethyl methacrylate) (PHEMA) beads were investigated for specific adsorption of yeast invertase from aqueous solutions. PHEMA beads were prepared by a suspension polymerization technique with an average size of 150-200 microm, and activated by epichlorohydrin. Con A was then immobilized by covalent binding onto these beads. The maximum Con A immobilization was found to be 10 mg/g. The invertase-loading capability of the PHEMA/Con A beads was 107 mg/g. The maximum invertase adsorption capacity on the PHEMA/Con A adsorbents was observed at pH 5.0. The values of the Michaelis constant K(m) of invertase were significantly larger upon adsorption, indicating decreased affinity by the enzyme for its substrate, whereas V(max) was smaller for the adsorbed invertase. Adsorption improved the pH stability of the enzyme as well as its temperature stability. Thermal stability was found to increase with adsorption. The adsorbed enzyme activity was found to be quite stable in repeated experiments. Storage stability of adsorbed invertase.

摘要

研究了用伴刀豆球蛋白A(Con A)固定化的聚甲基丙烯酸2-羟乙酯(PHEMA)微珠从水溶液中特异性吸附酵母转化酶的性能。PHEMA微珠通过悬浮聚合技术制备,平均粒径为150 - 200微米,并用环氧氯丙烷进行活化。然后通过共价结合将Con A固定在这些微珠上。发现Con A的最大固定量为10 mg/g。PHEMA/Con A微珠的转化酶负载能力为107 mg/g。在pH 5.0时观察到PHEMA/Con A吸附剂上转化酶的最大吸附容量。吸附后转化酶的米氏常数K(m)值显著增大,表明酶对其底物的亲和力降低,而吸附的转化酶的V(max)较小。吸附提高了酶的pH稳定性及其温度稳定性。发现热稳定性随吸附而增加。在重复实验中发现吸附的酶活性相当稳定。吸附的转化酶的储存稳定性。

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