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University of Iowa Research Foundation

Biotechnology Research and Development Corporation

Vikram M Paradkar

Jonathan S Dordick

Maria Vladimir Sergeeva

A method is presented for catalyzing the conversion of substrate into product in an organic reaction solvent with an enzyme-surfactant ion pair. The enzyme-surfactant ion pair comprises the enzyme catalyzing the reaction and a surfactant capable of forming an ion pair with the enzyme. Water present in the organic solvent at a concentration of about 0.03% to about 2.5% is sufficient to enhance the rate of catalysis and stabilize the enzyme without substantially increasing the rate of hydrolysis when compared to the anhydrous enzyme.

Enzyme catalysis in organic solutions containing water

Organic enzyme solutions comprise an ion pair complex of an enzyme and a surfactant, and an organic solvent in which the enzyme-surfactant ion pair is dissolved. The solution has catalytic activity at least an order of magnitude greater than a suspension of an equal amount of the enzyme and organic solvent without ion pair complexes. The enzyme is preferably a hydrolase with an acyl transferase activity and the surfactant is Aerosol OT (sodium bis(2-ethylhexyl) sulfosuccinate) at low concentrations. The organic enzyme solution may be made by obtaining an aqueous solution of an enzyme at a pH of maximal enzyme activity, adding an ionic surfactant to the aqueous solution in an amount sufficient to form an ion pair, and extracting the enzyme-surfactant ion pair into an organic solvent phase without substantial formation of reversed micelles, to produce an anhydrous organic enzyme solution comprising the enzyme-surfactant ion pair dissolved in the organic solvent such that the enzyme retains native structure and substantial catalytic activity. Catalytic solutions according to the invention can be used to catalyze peptide synthesis from acyl donors and nucleophiles.

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Madison, WI, United States of America

Vikram M Paradkar