Immobilization is a unique method for the improvement of product yield. During solid phase catalysis, the chemistry of the synthetic matrix plays an essential role in the performance of the biocatalyst. In the current study, immobilized protease within polyacrylamide macrosphere beads exhibited 76.0% entrapment yield with a remarkable stability of enzyme (33.0%) after 30 days of storage period at 4 °C. The entrapment of free enzyme within polyacrylamide also increased the optimal reaction temperature by 55 °C and provided a broad range of pH optima. Moreover, a significant enhancement in thermal stability was also detected. Polyacrylamide entrapped protease revealed up to 30.93% activity after incubation period of 30.0 min at 70 °C whereas, the free enzyme was completely inactivated at this temperature. Additionally, entrapped protease displayed an efficient recycling capacity and retained approximately 24.0% of its initial activity after eight successive reaction cycles. After entrapment of protease, the anchoring of substrate to the active site of the free protease exhibited change in K_m and V_max values. Therefore, owing to economic feasibility, the polyacrylamide entrapped protease might be a promising candidate for various applications in different industrial sectors.

固定化是提高产品产量的独特方法。在固相催化过程中，合成基质的化学性质在生物催化剂的性能中起着至关重要的作用。在当前的研究中，在4°C下储存30天后，固定在聚丙烯酰胺大球中的蛋白酶表现出76.0％的包封率，并具有显着的酶稳定性（33.0％）。聚丙烯酰胺中游离酶的截留也将最佳反应温度提高了55°C，并提供了最宽的pH最佳范围。此外，还发现了热稳定性的显着提高。聚丙烯酰胺捕获的蛋白酶在70°C下孵育30.0分钟后显示高达30.93％的活性，而游离酶在该温度下被完全灭活。此外，捕获的蛋白酶显示出有效的再循环能力，并且在八个连续的反应循环后保留了其初始活性的约24.0％。截留蛋白酶后，底物锚定在游离蛋白酶的活性位点上表现出变化。K _m和V _max值。因此，由于经济上的可行性，聚丙烯酰胺包埋的蛋白酶可能是在不同工业领域中的各种应用的有前途的候选者。