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water-immiscible organic solvents as in the dry powder, but the half-life of dilute solutions is very short [64]. Subtilisin crystalsplaced in acetonitrile have the same crystal structure as does subtilisin crystallized from aqueous solutions [41]. Enzymes do notturn inside-out when placed in immiscible organic solvents.The stability and catalytic activity of enzymes in water/water-miscible organic solvent systems are different from those in waterimmiscibleorganic solvent systems. Kang et al. [55–57] showed that protease-catalyzed hydrolysis of casein, in either 5%ethanol/95%aqueous buffer or 5% acetonitrile/95% aqueous buffer systems, caused an increase inKm,a decrease inVmax,andadecrease in stability (determined by circular dichroism and differential scanning calorimetry methods) compared to controls inaqueousbuffer only. It is well known that protic solvents, such as alcohols and amines, compete with water in hydrolytic enzymereactions[5, 9].7.5.6 Why Enzymes are Effective CatalystsEnzymes are very effective catalysts as indicated by their ability to lower the activation energy,Ea, of reactions (Table 2). Thisresults in larger values ofno. But these facts provide no indication of how the enzymes achieve such “magic.” Be assured thatenzymes act by the same principles of all chemical reactions. They just do it better [67].
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