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用于过氧化氢酶固定化的肟功能化的水凝胶盘。

Oxime-functionalized cryogel disks for catalase immobilization.

机构信息

Aksaray University, Technical Vocational School of Higher Education, Department of Chemistry and Chemical Processing Technology, Aksaray, Turkey.

Dokuz Eylul University, Faculty of Science, Department of Chemistry, Izmir, Turkey.

出版信息

Int J Biol Macromol. 2018 Jul 15;114:812-820. doi: 10.1016/j.ijbiomac.2018.04.006. Epub 2018 Apr 5.

Abstract

Catalase is a protective enzyme against oxidative stress and converts hydrogen peroxide into water and molecular oxygen. In the current study, catalase immobilization was applied onto the oxime-functionalized cryogel disks. Cryogel disks were produced by free radical polymerization. After cutting as circular disks, oxime ligand (4-biphenylchloroglyoxime, BPCGO) was attached and oxime-functionalized cryogel disks were obtained. After optimization of several immobilization parameters such as pH, initial catalase concentration, temperature and ionic strength, maximum catalase load was detected as 261.7 ± 11.2mg/g for cryogel disk at pH5.0. Activity studies indicated that immobilization enhanced the enzyme activity in basic pH region, the temperature range of 15-35°C and at ionic strengths between 0.2 and 1.0M NaCl. Km was detected as 9.9 and 11.0mM and V was 357.1 and 769.2μmol min for free and immobilized catalase, respectively. k and Km/k values showed that immobilization enhanced the catalytic efficiency. Storage stability experiments demonstrated that immobilization increased the usability period. Furthermore, catalase desorption was achieved by 1.0M NaSCN at pH8.0 successfully and catalase adsorption capacity of oxime-functionalized cryogel disk was decreased by 9.9% at the end of 5 adsorption-desorption cycle.

摘要

过氧化氢酶是一种抵抗氧化应激的保护酶,可将过氧化氢转化为水和分子氧。在本研究中,将过氧化氢酶固定化应用于肟功能化的水凝胶盘上。水凝胶盘是通过自由基聚合制成的。切割成圆形盘后,连接肟配体(4-联苯氯代乙二肟,BPCGO),得到肟功能化的水凝胶盘。在优化了 pH、初始过氧化氢酶浓度、温度和离子强度等几个固定化参数后,在 pH5.0 时,水凝胶盘的最大过氧化氢酶负载量检测为 261.7±11.2mg/g。活性研究表明,固定化在碱性 pH 区域、15-35°C 的温度范围和 0.2-1.0M NaCl 的离子强度下增强了酶的活性。Km 分别为 9.9 和 11.0mM,V 分别为 357.1 和 769.2μmol min,游离和固定化过氧化氢酶。k 和 Km/k 值表明固定化提高了催化效率。储存稳定性实验表明,固定化增加了可用性期限。此外,通过在 pH8.0 下用 1.0M NaSCN 成功实现了过氧化氢酶的解吸,并且在 5 次吸附-解吸循环结束时,肟功能化水凝胶盘的过氧化氢酶吸附容量降低了 9.9%。

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