Formation of protein-based coatings and structures via Enzyme Mediated Autodeposition (EMA) is investigated on the example of casein as protein and chymosin as enzyme. Key factor of this method is the tethering of the enzyme onto the support. By this, destabilization and subsequent deposition of casein particles occur only in direct proximity to the support surface. Investigated immobilization methods are physical adsorption, direct covalent binding, and covalent binding via polymeric spacers.Physical adsorption of enzyme enables the formation of continuous casein coatings with controllable film thickness by adjustment of reaction parameters, such as deposition time, pH value, and particle concentration. This method is also suitable for the in situ buildup of adhesive protein layers. Highest control and site-specificity of the process is provided by covalent attachment of enzyme. Direct covalent tethering results in defined formation of protein mono- or double layers, while the incorporation of spacer molecules enhances the mobility of enzyme. Consequently, the amount and radius of protein deposition is increased. Combination of the EMA with the Nanosphere Lithography technique allows for tethering of enzyme only in designated areas. This enables the controlled deposition of single protein particles and allows for the nanostructuring of surfaces with biopolymers, providing an unrivaled precision and economic cost-efficiency.Based on the attained results, the Enzyme Mediated Autodeposition provides new insights into biobased material design and might find applications in biosensors, micro- and nanoelectronics, and life-sciences.