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固定化对聚(HEMA-GMA)冷冻凝胶携带的过氧化氢酶活性的影响。

Effect of immobilization on the activity of catalase carried by poly(HEMA-GMA) cryogels.

机构信息

Osmancik Omer Derindere Vocational School, Department of Property Protection and Safety, Hitit University, Corum 19500, Turkey.

Alaca Avni Celik Vocational School, Department of Food Processing, Hitit University, Corum 19600, Turkey.

出版信息

Int J Biol Macromol. 2019 Feb 15;123:738-743. doi: 10.1016/j.ijbiomac.2018.11.121. Epub 2018 Nov 16.

DOI:10.1016/j.ijbiomac.2018.11.121
PMID:30452980
Abstract

Hydrogen peroxide is converted by catalase to molecular oxygen and water to remove oxidative stress. In this study, catalase immobilization was performed using poly(2-hydroxyethyl methacrylate-glycidyl methacrylate) (poly(HEMA-GMA)) cryogels with different amounts of GMA. Catalase adsorption capacity of 298.7 ± 9.9 mg/g was achieved at the end of 9 h using the poly(HEMA-GMA)-250 cryogel. Kinetic parameters and the inhibitory effects of pesticides such as 4,4'-DDE and 4,4'-DDT on the activity of free and immobilized catalase enzyme were investigated. While the V value of the immobilized enzyme was reduced 4-fold compared to the free enzyme, in the case of the comparison of the K values, the affinity of the immobilized enzyme was increased by 1.94 times against the substrate. The inhibitory effect of 4,4'-DDT pesticide was found to be higher for the immobilized and free enzyme. NaCl (1 M, pH: 7.0) solution was used for desorption of the adsorbed catalase enzyme. A desorption ratio of 96.45% was achieved. The technique used in this study is promising regarding for the immobilization of catalase enzyme to increase the operational activity. Therefore, poly(HEMA-GMA) cryogels have the potential to be used for immobilization of catalase enzyme in the fields of biology and biochemistry.

摘要

过氧化氢在过氧化氢酶的作用下转化为分子氧和水,以去除氧化应激。在这项研究中,使用不同含量的 GMA 的聚(2-羟乙基甲基丙烯酸酯-甲基丙烯酸缩水甘油酯)(poly(HEMA-GMA))冷冻凝胶进行了过氧化氢酶的固定化。使用 poly(HEMA-GMA)-250 冷冻凝胶在 9 小时结束时实现了 298.7±9.9 mg/g 的过氧化氢酶吸附容量。研究了动力学参数和农药如 4,4'-DDE 和 4,4'-DDT 对游离和固定化过氧化氢酶活性的抑制作用。虽然固定化酶的 V 值比游离酶降低了 4 倍,但在比较 K 值的情况下,固定化酶对底物的亲和力增加了 1.94 倍。发现 4,4'-DDT 农药对固定化和游离酶的抑制作用更高。使用 1 M NaCl(pH:7.0)溶液进行吸附的过氧化氢酶的洗脱。实现了 96.45%的洗脱比。本研究中使用的技术在提高操作活性方面对过氧化氢酶的固定化具有广阔的前景。因此,聚(HEMA-GMA)冷冻凝胶具有在生物学和生物化学领域固定化过氧化氢酶的潜力。

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