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反应性绿 5 修饰的聚丙烯酰胺/壳聚糖水凝胶:一种用于过氧化氢酶的亲和基质。

Reactive Green 5-Decorated Polyacrylamide/Chitosan Cryogel: An Affinity Matrix for Catalase.

机构信息

Chemistry Department, Faculty of Arts and Sciences, Aydın Adnan Menderes University, 09010, Aydın, Turkey.

出版信息

Appl Biochem Biotechnol. 2020 Dec;192(4):1191-1206. doi: 10.1007/s12010-020-03393-9. Epub 2020 Jul 24.

Abstract

Acrylamide/chitosan-based cryogel was fabricated, and a triazine dye, Reactive Green 5, was attached to the cryogel by nucleophilic substitution to build a dye affinity support for adsorption of catalase enzyme. Characterization of cryogel was performed using FTIR, SEM, EDX, BET, and swelling test. Synthesized cryogel beared pores with ~ 200 μm in size and the surface area of 11.8 m/g. Maximum catalase adsorption was (17.6 ± 0.29 mg/g) measured at pH 4.0 and 25 °C. The adsorption sites on the cryogel were saturated at 0.75 mg/mL enzyme concentration. Increased ionic strength caused a decrease in adsorption capacity. Desorption of catalase from cryogel was enabled using 0.5 M NaSCN solution. Consecutive adsorption experiments were carried out fifteen times to evaluate the reusability of the cryogel. Thermal, storage, and operational stabilities of immobilized catalase were higher than the free one. The data produced implicate that catalase-adsorbed dye-affinity cryogel may be used for HO detection or removal when necessary. Graphical Abstract.

摘要

丙烯酰胺/壳聚糖基水凝胶被制备,三嗪染料活性绿 5 通过亲核取代反应连接到水凝胶上,构建了用于吸附过氧化氢酶的染料亲和支撑物。使用 FTIR、SEM、EDX、BET 和溶胀试验对水凝胶进行了表征。合成的水凝胶具有约 200 μm 的孔径和 11.8 m/g 的表面积。在 pH 4.0 和 25°C 下,测量到最大过氧化氢酶吸附量为 (17.6 ± 0.29 mg/g)。在 0.75 mg/mL 酶浓度下,水凝胶上的吸附位点达到饱和。离子强度增加导致吸附容量下降。0.5 M NaSCN 溶液可用于从水凝胶中解吸过氧化氢酶。进行了十五次连续吸附实验来评估水凝胶的可重复使用性。固定化过氧化氢酶的热、储存和操作稳定性均高于游离酶。所产生的数据表明,当需要时,吸附过氧化氢酶的染料亲和水凝胶可用于 HO 的检测或去除。

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