The current study describes the immobilization of β-galactosidase on glutaraldehyde modified chitosan coated manganese oxide nanoparticles (CS-MnO-NPs). A notably higher immobilization yield (89%) was obtained for the enzyme on the developed nanobiocatalyst. The activity of immobilized enzyme was significantly higher at varying pH and temperature ranges as compared to the free enzyme. It was observed that the immobilized enzyme retained 86% activity at pH 8.0 in contrast to 46% by the soluble enzyme under similar condition. Moreover, β-galactosidase retained 79% activity at 60°C as compared to 41% by the free enzyme under identical exposure. It was further noticed that the immobilized enzyme retained 63% activity at 150 mM galactose concentration. However, under similar experimental conditions, SβG showed 41% activity. Immobilized exhibited 86% activity even after its sixth repeated use. Furthermore, in comparison to the free enzyme, the enzyme conjugated to glutaraldehyde modified chitosan coated MnO-NPs showed pronounced conversion of lactose in batch reactors for extended periods of time at 40°C. Therefore, these nanotechnology-based platforms can facilitate the creation of biocatalysts based on the K. lactis β-galactosidase enzyme, with the goal of enhancing current bioprocesses.