Bioactive materials have been explored for a broad range of applications including biocatalysts, biosensors, antifouling membranes and other functional and smart materials. We report herein a unique method for preparation of bioactive materials through a spin coating process. Specifically, we investigated the preparation of protease Subtilisin Carlsberg-coated plastic films and examined their activities for hydrolysis of chicken egg albumin (CEA). The process generated enzymic coatings with a typical loading of 13 μg/cm2, retaining 46% of the enzyme activity for hydrolysis of CEA in aqueous solutions. Interestingly, the surface-coated protease thin film not only catalyzed the hydrolysis of CEA in aqueous solutions, but also showed good activity for solid-state CEA that was coated on top of the enzyme thin film.