Activating Biocatalysts for Nonaqueous and Combinatorial Reactions

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This proposal pursues an older observation that the lyophilization of certain enzyme catalysts in the presence of highly ionic, non-buffering salts, e.g., KCl, can increase the catalytic rate in non-aqueous systems by 3-4 orders of magnitude. This has been demonstrated primarily on subtilisin. The specific aims of the original proposal are: To elucidate the mechanism(s) of enzyme activation in organic solvents with saltsTo extend the concept to a broad range of enzymes and reactions. To utilize the activated enzymes for combinatorial biocatalysis. In the revised and funded proposal, Aim 3 has been dropped and a more detailed examination of the mechanism and scope of the phenomenon has been added.