We report on the encapsulation of a laccase solution in silica colloidosomes of different shell structures and the study of catalytic performance of the encapsulated enzyme in toluene. The encapsulation is carried out by linking silica nanoparticles at the water/oil interface using hyperbranched polyethoxysiloxane (PEOS) in an aqueous-laccase-solution-intoluene Pickering emulsion. By varying the weight ratio of silica particles to water (R-s/w), colloidosome shell can be adjusted from a particle monolayer up to a bilayer bound with a sandwiched nanoporous silica thin film formed from PEOS. The encapsulated laccase exhibits catalytic activity and reusability that are controlled by R-s/w. With the increase of R-s/w the reusability of the enzyme improves, meanwhile, its activity declines. This method allows fabricating enzyme microcapsules with tailored activity and controlled release properties.