Akdoğan Doruk, Peksel Ayşegül
Department of Chemistry, Faculty of Arts and Science, Yildiz Technical University, Istanbul, Turkey.
Department of Pharmacy Services, Health Services Vocational School, Istanbul Nisantasi University, Sariyer, Turkey.
Biotechnol Appl Biochem. 2025 Jun;72(3):652-663. doi: 10.1002/bab.2687. Epub 2024 Nov 3.
One of the main goals of contemporary biotechnology has been the development of novel immobilized enzyme formulations. In the present study, the industrially important β-galactosidase was trapped in a polyvinyl alcohol (PVA) gel to immobilize it. The optimization of immobilization method and characterization of the immobilized enzyme were studied. The results were compared with free enzymes. The results indicate that the optimal temperature range for the enzyme to be at following immobilization is between 40°C and 50°C. At pH 7, the optimal pH, the activity increased, the V value increased from 1.936 to 2.495 U mg, and the K value decreased from 4.861 to 0.982 mM. Depending on how stable the immobilized enzyme when stored, β-galactosidases immobilized on PVA gels showed 52.87% activity at the end of the seventh week and 58.86% activity at the end of the fifth week. Their initial activity subsided after three reuses. The final result was 66%. Therefore, one may argue that it increases the catalytic effect of the enzyme. As a result, it has been found that immobilized β-galactosidase has more potent enzymatic properties than free β-galactosidase, which may make it more advantageous for industrial processes. Further studies could delve deeper into the mechanistic aspects of the immobilization process in an effort to improve optimization and tailor the immobilized enzyme to specific industrial needs.
当代生物技术的主要目标之一是开发新型固定化酶制剂。在本研究中，具有工业重要性的β-半乳糖苷酶被包埋在聚乙烯醇（PVA）凝胶中以实现固定化。研究了固定化方法的优化以及固定化酶的表征，并将结果与游离酶进行了比较。结果表明，固定化后酶的最佳温度范围在40°C至50°C之间。在pH 7（最佳pH值）时，活性增加，V值从1.936 U mg增加到2.495 U mg，K值从4.861 mM降低到0.982 mM。根据固定化酶在储存时的稳定性，固定在PVA凝胶上的β-半乳糖苷酶在第七周结束时显示出52.87%的活性，在第五周结束时显示出58.86%的活性。经过三次重复使用后，它们的初始活性下降，最终结果为66%。因此，可以认为它提高了酶的催化效果。结果发现，固定化β-半乳糖苷酶比游离β-半乳糖苷酶具有更强的酶学性质，这可能使其在工业过程中更具优势。进一步的研究可以更深入地探讨固定化过程的机理方面，以努力改进优化并使固定化酶符合特定的工业需求。
