α-Galactosidases hold immense potential due to their biotechnological applications in various industrial and functional food sectors. In the present study, soluble and covalently cross-linked aggregated forms of a low molecular weight, thermo-labile α-galactosidase from Vigna mungo (VM-αGal) seeds were immobilized onto chitosan-coated magnetic nanoparticles for improved stability and repeated usage by magnetic separation. Parameters like precipitants (type, amount, and ratio), glutaraldehyde concentration, and enzyme load were optimized for the preparation of chitosan-coated magnetic nanocomposites of cross-linked VM-αGal (VM-αGal-MC) and VM-αGal (VM-αGal-M) resulted in 100% immobilization efficiency. Size and morphology of VM-αGal-M were studied through dynamic light scattering (DLS) and scanning electron microscopy (SEM), while Fourier transform infrared spectroscopy (FTIR) was used to study the chemical composition of VM-αGal-MC and VM-αGal-M. VM-αGal-MC and VM-αGal-M were found more active in a broad range of pH (3–8) and displayed optimal temperatures up to 25 °C higher than VM-αGal. Addition of non-ionic detergents (except Tween-40) improved VM-αGal-MC activity by up to 44% but negatively affected VM-αGal-M activity. Both VM-αGal-MC (15% residual activity after 21 min at 85 °C, E_d 92.42 kcal/mol) and VM-αGal-M (69.0% residual activity after 10 min at 75 °C, E_d 39.87 kcal/mol) showed remarkable thermal stability and repeatedly hydrolyzed the substrate for 10 cycles.

α-半乳糖苷酶由于在各种工业和功能性食品领域的生物技术应用而具有巨大的潜力。在本研究中，来自Vigna mungo的低分子量，热不稳定的α-半乳糖苷酶的可溶性和共价交联的聚集形式将（VM-αGal）种子固定在壳聚糖包被的磁性纳米颗粒上，以提高稳定性并通过磁分离重复使用。优化了沉淀剂（类型，含量和比例），戊二醛浓度和酶负荷等参数，以制备交联的VM-αGal（VM-αGal-MC）和VM-αGal（VM- αGal-M）导致100％的固定效率。通过动态光散射（DLS）和扫描电子显微镜（SEM）研究了VM-αGal-M的大小和形态，而傅立叶变换红外光谱（FTIR）用于研究VM-αGal-M和VM-的化学组成。 αGal-M。发现VM-αGal-MC和VM-αGal-M在广泛的pH值（3-8）中更具活性，并且显示的最佳温度比VM-αGal高25°C。添加非离子型去污剂（Tween-40除外）最多可将VM-αGal-MC活性提高44％，但对VM-αGal-M活性产生负面影响。两种VM-αGal-MC（85°C下21分钟后E残留活性为15％_d 92.42 kcal / mol）和VM-αGal-M（在75°C下放置10分钟后残留活性为69.0％，E _d 39.87 kcal / mol）表现出显着的热稳定性，并反复水解底物10个循环。