Abstract

Chitosan (CS) and glutaraldehyde (GA) were exploited to activate carrageenan-alginate (Car-Alg) beads and enable their utilisation as covalent immobilisation matrices. The GA/CS activated Car-Alg beads were utilised to immobilise the Aspergillus oryzae β-D-galactosidase (β-gal). Initially, the beads gelling solution was optimised, and it was shown that the upmost amount of immobilised β-gal (iβ-gal) was acquired upon utilising a 1% CaCl₂–2% KCl gelling solution. Afterwards, the GA/CS activation protocol was honed via the Box Behnken design (BBD). The BBD revealed that soaking the Car-Alg beads with a 1.62% CS solution of pH 6 followed by a 3% GA solution was the optimum activation protocol that would provide 13.94 U/g beads iβ-gal. Noteworthy, a 72.64% immobilisation yield was accomplished via the optimally activated Car-Alg beads. The free and the iβ-gals were compared with respect to their K_m and V_max values, their temperature profiles, and their pH profiles. The reusability of the iβ-gal was also assessed, and 79.74% of its first round activity was retained throughout the 10th reusability round.

抽象的

壳聚糖（CS）和戊二醛（GA）被用来激活角叉菜胶-藻酸盐（Car-Alg）珠，并使其可用作共价固定基质。GA / CS活化的Car-Alg珠用于固定米曲霉β-D-半乳糖苷酶（β-gal）。最初，对珠的胶凝溶液进行了优化，结果表明，使用1％CaCl ₂可获得最大量的固定化β-gal（iβ-gal）。–2％KCl胶凝溶液。之后，通过Box Behnken设计（BBD）完善了GA / CS激活协议。BBD揭示了用1.62％的pH 6的CS溶液和3％的GA溶液浸泡Car-Alg珠是最佳的激活方案，可以提供13.94 U / g珠iβ-gal。值得注意的是，通过最佳活化的Car-Alg珠实现了72.64％的固定化率。比较了游离的和iβ-gals的K _m和V _max值，它们的温度曲线和它们的pH曲线。还评估了iβ-gal的可重用性，并且在整个第十个可重用性回合中保留了其第一轮活动的79.74％。