Abstract Immobilization of enzymes from different sources on various supports in designed systems increases enzymes’ stability by protecting the active site of it from undesired effect of reaction environment. Also, immobilization decreases the cost of separation and facilities the reuse of the enzymes. Therefore, the design of new immobilization enzyme preparations has been an inevitable area of modern biotechnology. Herein, Rhizomucor miehei lipase (RML) was immobilized on montmorillonite K-10 (MMT-RML) by adsorption and in polyvinyl alcohol (PVA-RML) by entrapment to obtain a more stable and active lipase preparation. The free and immobilized lipase preparations were characterized for p-nitrophenyl palmitate hydrolysis. The apparent Michaelis–Menten (Kmapp) constant was almost the same for the free RML and PVA-RML, whereas the corresponding value was 17.7-fold lower for MMT-RML. PVA-RML and MMT-RML have shown a 1.1 and 23.8 folds higher catalytic efficiency, respectively, than that of the free RML. The half-lives of PVA-RML and MMT-RML were found to be 7.4 and 3.4 times longer than the free RML at 35 °C, respectively. PVA-RML and MMT-RML maintained 65% and 87% of their initial activities after four reuses. These results showed that the catalytic performance of RML has improved significantly by immobilization.

中文翻译：

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米黑根霉脂肪酶在蒙脱石 K-10 和聚乙烯醇凝胶上的固定化

摘要 在设计的系统中将不同来源的酶固定在各种支持物上，通过保护酶的活性位点免受反应环境的不利影响来增加酶的稳定性。此外，固定化降低了分离成本并便于酶的再利用。因此，设计新的固定化酶制剂已成为现代生物技术的必然领域。在此，米黑根霉脂肪酶 (RML) 通过吸附固定在蒙脱石 K-10 (MMT-RML) 上，并通过包埋固定在聚乙烯醇 (PVA-RML) 中，以获得更稳定和活性的脂肪酶制剂。游离的和固定化的脂肪酶制剂被表征为对硝基苯基棕榈酸酯水解。对于游离 RML 和 PVA-RML，表观 Michaelis-Menten (Kmapp) 常数几乎相同，而 MMT-RML 的相应值低 17.7 倍。PVA-RML 和 MMT-RML 的催化效率分别比游离 RML 高 1.1 和 23.8 倍。发现 PVA-RML 和 MMT-RML 在 35°C 下的半衰期分别比游离 RML 长 7.4 和 3.4 倍。在四次重复使用后，PVA-RML 和 MMT-RML 保持了其初始活性的 65% 和 87%。这些结果表明，通过固定化，RML 的催化性能得到了显着提高。在四次重复使用后，PVA-RML 和 MMT-RML 保持了其初始活性的 65% 和 87%。这些结果表明，通过固定化，RML 的催化性能得到了显着提高。在四次重复使用后，PVA-RML 和 MMT-RML 分别保持了 65% 和 87% 的初始活性。这些结果表明，通过固定化，RML 的催化性能得到了显着提高。