Biocatalysis has been well regarded as an important and cost-effective technology for the manufacture of fine chemicals, synthesis of pharmaceuticals, biosensors, bioremediation, protein digestion in proteomic analysis, and biofuel cells. Stability and activity are always the most concerned properties of enzymes and limited factors to the vary applications. Here we present a facile in-situ aqueous polymerization procedure to fabricate single enzyme nanogels as robust nanometer scale biocatalysis. By using versatile modification strategies, such as acryloylation of lysine group, introducing vinyl group to glutamic acid and asparagine, control of modification pH, active site protection by inhibitors, different single enzyme nanogels are successfully prepared, which show greatly improved stability to temperature, organic solvent and low pH solution without the compromise of catalytic capability. These nanogels are quite suitable to biocatalytic applications in harsh condition, which are constrainted by small size or high mass-transfer rating reaction or exposed for extreme pH and temperature.

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