Elucidation of recognition sites of lipase toward its development as chemical catalysis

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The aim of this project is to realize and control an enzymatic reaction as a chemical reaction. The research results are as follows ;(1) Candida rugosa lipase (CRL) was treated with several concentrations of 2-propanol in water and the changes of the amount of the recovered protein, the hydrolytic activity, and the enentioselectivity toward methyl 2-phenylpropanoate were studied. The results shows that the conformational change of CRL occurred in 30% 2-propanol treatment.(2) The sizes of the large, medium, and small substituent recognition sites (L, M, and S pocket, respectively) in CRL were roughly estimated by measuring the specific hydrolytic activity against several p-nitrophenyl esters. These relative sizes were assessed as L pocket > phenyl, ethyl > M pocket > methyl > S pocket > hydrogen.(3) The highly enantioselective transesterification of 2-phenyl-1-propanol, which has not been efficiently resolved by lipase-catalyzed reactions, was attained by using vinyl esters having aromatic ring in acyl moiety as vinyl 3-phenylpropanoate.