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Oriented assembly of protein on surfaces

Several approaches that facilitate the oriented assembly of proteins on a variety of surfaces, including membranes, is described. Biotinylation reactions performed under controlled conditions were used to immobilize enzymes in a layer-by-layer fashion onto surfaces modified with avidin or streptavidin. For other enzymes, a specific attachment site was introduced by gene fusion or site-directed mutagenesis. These strategies were applied to achieve site-specific immobilization of several enzymes including alkaline phosphatase, organophosphorus hydrolase, horseradish peroxidase, and subtilisin. The site-specific immobilization led to orientation of the enzyme molecules on the surface of materials and to a higher activity compared to conventional immobilization methods.