In this study, a novel way to immobilize the enzyme β-galactosidase (Kluyveromyces lactis) through complexation with polysaccharides (sodium alginate (ALG) or Ɩ-carrageenan (CA)) was described. The kinetic parameters of the formation of the complexes and the immobilization efficiency of the enzyme as a function of pH and ratio of biopolymers were studied. The temperature profile, thermal stability, and kinetic parameters of the immobilized and free lactase were also analyzed. At pH 4.0, the results for the zeta potential associated with complex yield showed that the highest complex yield (97.14% ± 2.14) was reached for LAC/ALG at a 5:1 ratio, whereas the highest complex yield (95.26% ± 2.44) was reached an 8:1 ratio for LAC/CA. The process of complexation decreased the activity retained by the enzyme after dissociation. However, this decrease was less substantial for the LAC/ALG (83.2 ± 3.7%) complex than for LAC/CA (69.4 ± 7.6%) complex. The Michaelis–Menten constant (K_m) increased from 2.91 mM (native enzyme) to 3.43 mM (LAC/ALG complex) and 4.15 (LAC/CA complex), while the apparent turnover number (K_cat) and the specificity constant (K_cat/K_m) of both systems were not significantly different. The immobilization of the enzyme by complexation did not increase its thermal stability. However, the activity of the complexed lactase remained higher than that of native enzyme when exposed to low pH (pH 4.0) after 200 and 250 min for the LAC/ALG and LAC/CA complexes respectively. Our findings suggest that complexation can be a useful method for the immobilization of β-galactosidase.

在这项研究中，一种固定酶β-半乳糖苷酶（乳酸克鲁维酵母）的新方法）通过与多糖（藻酸钠（ALG）或β-角叉菜胶（CA））的络合进行了描述。研究了配合物形成的动力学参数和酶的固定化效率与pH和生物聚合物比例的关系。还分析了固定和游离乳糖酶的温度曲线，热稳定性和动力学参数。在pH 4.0时，与复合物产率相关的ζ电位的结果表明，LAC / ALG以5：1的比例达到了最高复合物产率（97.14％±2.14），而最高复合物产率（95.26％±2.44） LAC / CA的比例达到8：1。络合过程降低了酶解离后保留的活性。但是，与LAC / CA（69.4±7）相比，LAC / ALG（83.2±3.7％）复合物的下降幅度较小。6％）复杂。米氏常数-（Mentis constant）（K _m）从2.91 mM（天然酶）增加到3.43 mM（LAC / ALG复合物）和4.15（LAC / CA复合物），而表观转换数（K _cat）和特异性常数（K _cat / K _m）两种系统均无显着差异。通过络合固定化酶不会增加其热稳定性。然而，分别对LAC / ALG和LAC / CA复合物暴露200和250分钟后，暴露于低pH（pH 4.0）时，复合乳糖酶的活性仍高于天然酶。我们的发现表明，络合可以是固定β-半乳糖苷酶的有用方法。