Development of a hybrid bioinorganic nanobiocatalyst: Remarkable impact of the immobilization conditions on activity and stability of β‐galactosidase

Hybrid bioinorganic biocatalysts have received much attention due to their simple syn-thesis, high efficiency, and structural features that favor enzyme activity and stability. The present work introduces a biomineralization strategy for the formation of hybrid nanocrystals from β‐ga-lactosidase. The effects of the immobilization conditions were studied, identifying the important effect of metal ions and pH on the immobilization yield and the recovered activity. For a deeper understanding of the biomineralization process, an in silico study was carried out to identify the ion binding sites at the different conditions. The selected β‐galactosidase nanocrystals showed high specific activity (35,000 IU/g biocatalyst) and remarkable thermal stability with a half‐life 11 times higher than the soluble enzyme. The nanobiocatalyst was successfully tested for the synthesis of galacto‐oligosaccharides, achieving an outstanding performance, showing no signs of diffusional limitations. Thus, a new, simple, biocompatible and inexpensive nanobiocatalyst was produced with high enzyme recovery (82%), exhibiting high specific activity and high stability, with promis-ing industrial applications.