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将酪氨酸酶固定在改性硅藻生物硅上:从水溶液中酶法去除酚类化合物。

Immobilization of tyrosinase on modified diatom biosilica: enzymatic removal of phenolic compounds from aqueous solution.

机构信息

Biochemical Processing and Biomaterial Research Laboratory, Faculty of Science, Gazi University, 06500 Teknikokullar, Ankara, Turkey.

出版信息

J Hazard Mater. 2013 Jan 15;244-245:528-36. doi: 10.1016/j.jhazmat.2012.10.041. Epub 2012 Oct 30.

Abstract

Acid and plasma treated diatom-biosilica particles, were modified with 3-aminopropyl triethoxysilane (APTES), and activated with glutaraldehyde. Then, tyrosinase was immobilized onto the pre-activated biosilica by covalent bonding. The biosilica properties were determined using SEM, and FTIR. The enzyme system has been characterized as a function of pH, temperature and substrate concentration. Optimum pH of the free and immobilized enzyme was found to be pH 7.0. Optimum temperatures of the free and immobilized enzymes were determined as 35 and 45 °C respectively. The biodegradation of phenolic compounds (i.e., phenol, para-cresol and phenyl acetate) has been studied by means of immobilized tyrosinase in a batch system. The immobilized tyrosinase retained about 74% of its original activity after 10 times repeated use in the batch system. Moreover, the storage stability of the tyrosinase-biosilica system resulted excellent, since they maintained more than 67% of the initial activity after eighth week storage. Highly porous structure of biosilica can provide large surface area for immobilization of high quantity enzyme. The porous structure of the biosilica can decrease diffusion limitation both substrate phenols and their products. Finally, the immobilized tyrosinase was used in a batch system for degradation of three different phenols.

摘要

酸和等离子体处理的硅藻生物硅颗粒,用 3-氨丙基三乙氧基硅烷(APTES)进行改性,并用过氧化戊二醛进行活化。然后,通过共价键将酪氨酸酶固定在预活化的生物硅上。使用 SEM 和 FTIR 确定生物硅的性质。研究了酶系统作为 pH、温度和底物浓度函数的特性。游离酶和固定化酶的最佳 pH 值均为 7.0。游离酶和固定化酶的最佳温度分别确定为 35 和 45°C。通过固定化酪氨酸酶在批处理系统中研究了酚类化合物(即苯酚、对甲酚和苯乙酸酯)的生物降解。固定化酪氨酸酶在批处理系统中重复使用 10 次后保留了其原始活性的约 74%。此外,酪氨酸酶-生物硅系统的储存稳定性非常出色,因为它们在第八周储存后仍保持初始活性的 67%以上。生物硅的高度多孔结构可为固定化大量酶提供大的表面积。生物硅的多孔结构可以减少底物苯酚及其产物的扩散限制。最后,将固定化酪氨酸酶用于批处理系统中,以降解三种不同的苯酚。

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