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Enzyme immobilization usually make use of nanomaterials to hold up biocatalysis stability in various unamiable reaction conditions, but also lead large discount on enzyme activity. Thus, there are abundant researches focus on how to deal with the relation of enzyme molecules and supports. In this work, a new state of highly active enzymes has been established through facile and novel in situ immobilization and soft template removal method to construct enzyme contained hollow silica nanosphere (catalase@HSN) biocatalysts where enzymes in the cavity exhibit "immobilized but not rigid state". The obtained catalase@HSN was characterized by transmission electron microscopy, scanning electron microscopy and confocal laser scanning microscopy et al. Catalase@HSN exhibits excellent activity (about 80 % activity recovery rate) and stability suffers from extreme pH, temperature, and organic solvents. Moreover, the reusability and storage stability of catalase@HSN also are satisfactory. This proposed strategy provides a facile method for preparing biocatalysts under mild conditions, facilitating the applications of immobilized enzyme in the fields of real biocatalytic industry with high apparent activity and passable stability. A new state of highly active enzymes has been established through soft template removal method to construct enzyme contained hollow silica nanosphere (enzyme@HSN) biocatalysts where enzymes in the cavity exhibit "immobilized but not rigid state". [Display omitted] • Hybrid MOF using glutathione as alternative ligand was used as self-sacrificial template for hollow silica fabrication. • Enzyme molecules in hollow silica nanosphere are "Immobilized but Not Rigid State" with improved activity. • This proposed strategy provides a facile method for preparing nano-biocatalysts under mild conditions. [ABSTRACT FROM AUTHOR]