Stability of hydrolytic enzymes in water-organic solvent systems |
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| Institution: | 1. Department of Biochemistry, József Attila University, P.O. Box 533, H-6701 Szeged, Hungary;2. COVENT Industrial Venture Capital Investment Co., Ltd., P.O. Box 335, H-1537 Budapest, Hungary;1. Department of Agricultural & Food Engineering, Indian Institute of Technology, Kharagpur 721302, India;2. Department of Organic Chemistry, Indian Institute of Science, Bangalore 560012, India;1. A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky pr. 33, bld. 2, 119071, Moscow, Russian Federation;2. Institute of Mathematical Problems of Biology, RAS, Institutskaya str. 4, Pushchino, 142290, Russian Federation;3. M.K. Ammosov North-Eastern Federal University, Belinskiy str., 58, Suite 312, Yakutsk, 677980, Republic of Sakha (Yakutia), Russian Federation;4. Dep. “Protein Factory”, NBICS Center, National Research Centre “Kurchatov Institute”, Akad. Kurchatova sqr., 1, Moscow, 123182, Russian Federation;1. College of Physics and Electronic Information Engineering, Wenzhou University, Wenzhou 325035, China;2. Department of Chemical Engineering, National Tsing Hua University, Hsinchu 30013, Taiwan;1. Departamento de Ingeniería Química, Facultad de Ciencias y Tecnologías Químicas, Universidad de Castilla-La Mancha, Avenida Camilo José Cela 12, 13005 Ciudad Real, Spain;2. Université de Lorraine, Institut Jean Lamour, UMR7198, F-54011 Nancy, France;3. CNRS, Institut Jean Lamour, UMR7198, F-54011 Nancy, France |
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| Abstract: | The effects of organic solvents on the stabilities of bovine pancreas trypsin, chymotrypsin, carboxypeptidase A and porcine pancreas lipase were studied. Water-miscible solvents (ethanol, acetonitrile, 1,4-dioxane and dimethyl sulfoxide) and water-immiscible solvents (ethyl acetate and toluene) were used in 100 mM phosphate buffer (pH 7.0) or 100 mM Tris/HCl buffer (pH 7.0) in concentrations of 20–80% (v/v). All hydrolytic enzymes studied were inactivated by mixtures containing dimethyl sulfoxide at higher concentrations. Trypsin and carboxypeptidase A resisted solvent mixtures containing acetonitrile, 1,4-dioxane and ethanol. They preserved more than 80% of their starting activities during 20-min incubations. The activities of lipase and chymotrypsin decreased with increasing concentration of water-miscible polar organic solvents, but at higher concentrations (80%) 70–90% of the activity remained. In mixtures with water-immiscible solvents, the decrease in activity of carboxypeptidase A was pronounced. Trypsin and chymotrypsin underwent practically no loss in activity in the presence of toluene or ethyl acetate. In respect of stability, the polar solvent proved to be more favorable for lipase. These results suggest that the conformational stabilities of hydrolytic enzymes are highly dependent on the solvent-protein interactions and the enzyme structure. |
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