Polyethyleneimine (PEI) glutaraldehyde-refined calcium pectinate (CaP)-agar beads were presented as improved covalent immobilization matrices. The CaP-agar beads exhibited incremented mechanical stability which facilitated their handling. The beads’ concoction and activation processes were honed using the Box–Behnken design which recommended utilizing 5.4% agar, and a 2.95% PEI solution of pH 8.67. The honed CaP-agar beads established a more efficient ionic interaction with PEI which enabled the immobilization of more enzyme while utilizing less PEI than that required to activate the neat CaP beads. Furthermore, the activated CaP-agar beads granted superior operational stability to the immobilized enzyme, β-d-galactosidase (βgal), where it preserved 86.84 ± 0.37% of its precursive activity during its thirteenth reusability round. The CaP-agar immobilized βgal (iβgal) also showed incremented storage stability where it preserved 85.05 ± 3.32% of its precursive activity after 38 days of storage. The thermal stability of the iβgal was shown to be superior to that of the free enzyme as the iβgal exhibited incremented thermodynamic parameters, such as the t_1/2 values, the D values, the thermal denaturation activation energy, the enthalpies, and the Gibb’s free energies. The βgal’s immobilization onto the activated CaP-agar beads also shifted the enzyme’s optimal pH from 4.6–5.1 to 3.3–4.9, whereas its optimal temperature was retained at 55 °C. The procured biocatalyst was exploited to efficiently hydrolyze the lactose in whey permeate.

聚乙烯亚胺 (PEI) 戊二醛精制果胶酸钙 (CaP)-琼脂珠作为改进的共价固定基质呈现。CaP-琼脂珠表现出增加的机械稳定性，这有利于它们的处理。珠子的混合和活化过程使用 Box-Behnken 设计进行磨练，该设计建议使用 5.4% 的琼脂和 2.95% 的 pH 8.67 的 PEI 溶液。磨光的 CaP-琼脂珠与 PEI 建立了更有效的离子相互作用，与激活纯 CaP 珠所需的 PEI 相比，这能够固定更多的酶，同时使用更少的 PEI。此外，活化的 CaP-琼脂珠赋予固定化酶 β- d优异的操作稳定性-半乳糖苷酶 (βgal)，在第十三轮可重复使用期间，它保留了 86.84 ± 0.37% 的前驱活性。CaP-agar 固定化 βgal (iβgal) 也显示出增加的储存稳定性，在储存 38 天后它保留了其前体活性的 85.05 ± 3.32%。iβgal 的热稳定性显示出优于游离酶的热稳定性，因为 iβgal 表现出增加的热力学参数，例如t _1/2值、D值、热变性活化能、焓和吉布斯自由能。βgal 固定在活化的 CaP-琼脂珠上也使酶的最佳 pH 值从 4.6-5.1 变为 3.3-4.9，而其最佳温度保持在 55°C。采购的生物催化剂用于有效水解乳清渗透物中的乳糖。