Distinct effect of a cationic surfactant on the transient and steady state phases of 2-naphthyl acetate hydrolysis catalyzed by α-chymotrypsin

Results obtained on the effect of addition of dodecyltrimethylammonium bromide (DTAB) upon the alpha-chymotrypsin (alpha-CT) catalyzed hydrolysis of 2-naphthyl acetate (2-NA) under steady state conditions for the acyl-enzyme intermediate are compared with those previously obtained in the transient (pre-steady state or "burst") phase. It is found that, while in the transient phase there is no effect of DTAB addition on the kinetic parameters at concentrations below the critical micelle concentration (CMC) of the surfactant, super-activity is observed when the acyl-enzyme intermediate reaches the steady state condition. This difference implies that the surfactant does not modify either the formation or the decomposition of the enzyme-substrate complex (transient phase) but notably increases the rate of disruption of the acyl-enzyme intermediate. (C) 2004 Elsevier B.V. All rights reserved.