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辣根过氧化物酶通过功能化 RGO-SiO 纳米复合材料共价固定化对苯酚和染料的生物降解。

Biodegradation of phenol and dyes with horseradish peroxidase covalently immobilized on functionalized RGO-SiO nanocomposite.

机构信息

Young Researchers and Elite Club, Shahr-e-Qods Branch, Islamic Azad University, Tehran, Iran.

Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran; Center of Excellence in Biothermodynamics, University of Tehran, Tehran, Iran.

出版信息

Int J Biol Macromol. 2020 Dec 1;164:4403-4414. doi: 10.1016/j.ijbiomac.2020.09.045. Epub 2020 Sep 12.

Abstract

Horseradish peroxidase (HRP) was immobilized onto a functionalized reduced graphene oxide-SiO through the covalent bonding process. By using scanning electron microscopy (SEM) and Fourier transform infrared (FT-IR), the formed nanocomposites were characterized. The kinetic parameters including the catalytic constant, k, and the catalytic efficiency, k/K, increased 5.5 and 6 times, respectively, after immobilization. The circular dichroism analysis demonstrated that the α-helical content increased from 39% to 46% after immobilization. The immobilization improved the reusability of HRP as 70% of initial activity retained after 10 cycles. Due to the buffering effect, the immobilized HRP was less sensitive to pH changes as compared to the free HRP. At temperature 40 °C and during 90 min, the immobilized HRP retained 90% of the initial activity while 70% of initial activity remained for the free HRP. After 35-day storage, no reduction in the activity was observed for the immobilized HRP. The removal efficiency for phenol concentration (2500 mg/L) obtained 100% and 50% for the immobilized and free HRP, respectively. The results showed that the immobilized HRP promoted the dyes decolorization from 2-fold until 26-fold as compared to the free HRP. The decolorization efficiencies reached 100% for most dyes in the case of immobilized HRP.

摘要

辣根过氧化物酶(HRP)通过共价键过程固定在功能化的还原氧化石墨烯-SiO 上。通过扫描电子显微镜(SEM)和傅里叶变换红外(FT-IR)对形成的纳米复合材料进行了表征。动力学参数包括催化常数 k 和催化效率 k/K,固定化后分别增加了 5.5 倍和 6 倍。圆二色分析表明,固定化后α-螺旋含量从 39%增加到 46%。固定化提高了 HRP 的可重复使用性,10 次循环后保留了初始活性的 70%。由于缓冲效应,与游离 HRP 相比,固定化 HRP 对 pH 变化的敏感性较低。在 40°C 温度下和 90 分钟内,固定化 HRP 保留了初始活性的 90%,而游离 HRP 保留了初始活性的 70%。在 35 天储存后,固定化 HRP 的活性没有下降。对于 2500mg/L 的苯酚浓度,固定化和游离 HRP 的去除效率分别为 100%和 50%。结果表明,与游离 HRP 相比,固定化 HRP 可将染料脱色效率提高 2 倍至 26 倍。对于固定化 HRP,大多数染料的脱色效率达到 100%。

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