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Abstract：
In the presented study, a variety of hybrid and single nanomaterials of various origins were tested as novel platforms for horseradish peroxidase immobilization. A thorough characterization was performed to establish the suitability of the support materials for immobilization, as well as the activity and stability retention of the biocatalysts, which were analyzed and discussed. The physicochemical characterization of the obtained systems proved successful enzyme deposition on all the presented materials. The immobilization of horseradish peroxidase on all the tested supports occurred with an efficiency above 70%. However, for multi-walled carbon nanotubes and hybrids made of chitosan, magnetic nanoparticles, and selenium ions, it reached up to 90%. For these materials, the immobilization yield exceeded 80%, resulting in high amounts of immobilized enzymes. The produced system showed the same optimal pH and temperature conditions as free enzymes; however, over a wider range of conditions, the immobilized enzymes showed activity of over 50%. Finally, a reusability study and storage stability tests showed that horseradish peroxidase immobilized on a hybrid made of chitosan, magnetic nanoparticles, and selenium ions retained around 80% of its initial activity after 10 repeated catalytic cycles and after 20 days of storage. Of all the tested materials, the most favorable for immobilization was the above-mentioned chitosan-based hybrid material. The selenium additive present in the discussed material gives it supplementary properties that increase the immobilization yield of the enzyme and improve enzyme stability. The obtained results confirm the applicability of these nanomaterials as useful platforms for enzyme immobilization in the contemplation of the structural stability of an enzyme and the high catalytic activity of fabricated biocatalysts.
摘要：
在提出的研究中，测试了各种来源的各种杂种和单一纳米材料作为辣根过氧化物酶固定化的新型平台。进行了彻底的表征，以建立固定的支持材料的适用性，以及生物催化剂的活性和稳定性保留，进行了分析和讨论。获得的系统的物理化学表征证明了酶在所有提供的材料上的成功沉积。辣根过氧化物酶在所有测试支持物上的固定效率高于70％。然而,对于多壁碳纳米管和由壳聚糖制成的杂化物，磁性纳米粒子，和硒离子，达到90%。对于这些材料,固定化产率超过80%，导致大量的固定化酶。产生的系统显示出与游离酶相同的最佳pH和温度条件；但是，在更广泛的条件下，固定化酶活性超过50%。最后,可重复使用性研究和储存稳定性测试表明，辣根过氧化物酶固定在壳聚糖杂种上，磁性纳米粒子，在10个重复的催化循环和20天的储存后，硒离子保留了其初始活性的80％左右。在所有测试材料中，最有利于固定化的是上述壳聚糖基杂化材料。所讨论的材料中存在的硒添加剂赋予其补充性质,所述补充性质增加酶的固定化产率并改善酶稳定性。获得的结果证实了这些纳米材料在酶的结构稳定性和制造的生物催化剂的高催化活性方面作为酶固定化的有用平台的适用性。
