In the current work, electrostatic interactions were used to immobilize the horseradish peroxidase (HRP) onto five types of ceramic materials (C) with different concentrations of oxidized metals (C1-C5). The highest immobilization efficiency (70 and 77%) was detected at 6 mg C3 and 18 enzyme units. Scanning Electron Microscope (SEM), Energy Dispersive X-ray (EDX) and Fourier Transform Infrared (FTIR) analysis of C3-HRP confirmed the immobilization of the enzyme. After ten reuses, the reusability analysis showed that (66%) of the C3-HRP enzyme activity was retained. For C3-HRP, the optimum pH and temperature of the soluble enzyme were shifted from 7.0 and 30 °C to 6.0 and 50 °C. Up to 40 °C and 50 °C, respectively, the soluble HRP and C3-HRP remained steady. The kinetic analysis revealed that the Km and Vmax of soluble HRP and C3-HRP were, respectively, 5.5 mM, 0.66 units, and 8 mM, 0.52 units for hydrogen peroxide (H₂O₂) and 35.5 mM, 3.4 units and 40 mM, 1.1 units for guaiacol. Compared to soluble-HRP, the C3-HRP exhibited a greater oxidizing affinity toward several phenolic compounds (Guaiacol, o-dianisidine, o-phenylenediamine, pyrogallol, p-aminoantipyrine). In comparison with soluble-HRP, the C3-HRP showed increased stress tolerance with Triton X-100, urea, metals, isopropanol, and dimethyl sulfoxide. The C3-HRP removed methyl orange more effectively compared to soluble-HRP.

Keywords: Ceramics; Characterization; Decolorization; Horseradish peroxidase; Immobilization.

© 2024. The Author(s).

在当前研究中，使用静电相互作用将过氧化物酶（HRP）固定到五种不同氧化金属浓度的陶瓷材料（C1-C5）上。在6毫克C3和18个酶单位的情况下检测到了最高的固定效率（70%和77%）。扫描电子显微镜（SEM）、能量散射X射线（EDX）和傅里叶变换红外光谱（FTIR）分析确认了C3-HRP中的酶固定。经过十次重复使用后，可重复使用的分析表明C3-HRP的酶活性保留率为66%。对于C3-HRP而言，溶解酶的最佳pH值和温度从7.0和30°C分别转移到6.0和50°C。在40°C和50°C时，溶解HRP和C3-HRP保持稳定状态。动力学分析表明，溶解HRP和C3-HRP对过氧化氢（H₂O₂）的Km值分别为5.5 mM和8 mM，Vmax值分别为0.66单位和0.52单位；对愈创木酚的Km值分别为35.5 mM和40 mM，Vmax值分别为3.4单位和1.1单位。与溶解HRP相比，C3-HRP表现出更强的氧化亲合力以处理多种酚类化合物（如愈创木酚、邻二甲苯基、对苯二胺、没食子酸、对氨基抗坏血酸）。相比于溶解HRP，C3-HRP显示出更高的压力耐受性，在Trition X-100、尿素、金属离子、异丙醇和二甲基亚砜中的表现更佳。C3-HRP比溶解HRP更为有效地去除品红。

关键词：陶瓷；表征；脱色；辣根过氧化物酶；固定化。

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