Nα-Boc-L-lysine 7-amido-4-methylcoumarin acetate salt

Formation of wormlike micelles in mixed anionic/cationic system without the addition of any salt has been studied. Amino-acid based anionic surfactant N-dodecylglutamic acid (LAD), which is practically immiscible with water at 25 degrees C upon neutralization by 2,2',2''-nitrilotriethanol (TEA) forms small micellar aggregates and the solution behaves like a Newtonian fluid. The rheological behavior of LAD/water/hexadecyltrimethylammonium bromide (CTAB) and LAD/water/dodecyltrimethylammonium bromide (DTAB) systems were investigated at different degrees of neutralization of the LAD depending on the concentration of the cationic surfactants and on temperature. Addition of CTAB to the dilute aqueous solution of the LAD-TEA-x (the neutralized product, where x represents the mole ratio of TEA) causes one dimensional micellar growth. After certain concentration the elongated micelles entangle forming a rigid network of viscoelastic wormlike micelles. Thus formed viscoelastic solutions follow Maxwellian behavior over a wide range of frequency and thus are considered to consist of transient network of wormlike micelles. By varying the degree of neutralization from 1:1 via 1:1.5 to 1:2 (molar ratio) phase and rheological behavior were modified in that the highly viscous region of viscoelastic wormlike micelles shifted to higher CTAB concentrations and no maxima in the zero-shear viscosity could be observed for the higher degree of neutralization of the LAD (1:1.5 and 1:2). However, the obtained rheological parameters showed scaling relationships that were consistent with the living polymer model. The zero-shear viscosity decays exponentially with temperature following Arrhenius behavior. The flow activation energy calculated from the Arrhenius plot is very close to the value reported for the typical wormlike micellar solution. In contrast to CTAB no formation of viscoelastic wormlike micelles could be observed with DTAB, although, the solution viscosity increases. The elongated micelles could not entangle to form a rigid network of wormlike micelles in this system. For the first time viscoelastic wormlike micelles could be obtained in salt-free mixed anionic/cationic surfactant systems.

The rate of hydrolysis of N-glutaryl-L-phenylalanine p-nitroanilide (GPNA) catalyzed by alpha-chymotrypsin (alpha-CT) has been measured in aqueous solutions of dodecyltrimethylammonium bromide (DTAB) at concentrations below and above the critical micelle concentration, as well as in the absence of surfactant. Under all the conditions employed, the reaction follows a Michaelis-Menten mechanism. The presence of the surfactant leads to superactivity below and above the critical micelle concentration (CMC), with a maximum reaction rate taking place near the CMC when the results are treated in terms of the analytical concentration of the substrate. A similar behavior was observed by working with the enzyme partially deactivated in the presence of 4 M urea. After correction to take into account the partitioning of the substrate between the micelles and the external media, the activity of the enzyme tends to remain almost constant above the corresponding CMCs. This results from a compensation of a decrease in the catalytic constant (k(cat)) and a decrease in the Michaelis constant (K(M)). The behavior of alpha-CT in the hydrolysis of GPNA in DTAB solutions is at variance with that previously reported for the hydrolysis of 2-naphthyl acetate in solutions of the same surfactant (E. Abuin, E. Lissi, R. Duarte, Langmuir 19 (2003) 5374). An explanation of the different effects of the surfactant on the behavior of the enzyme with both substrates is advanced, taking into account the complexity of the mechanism of the alpha-CT-mediated reaction, more specifically, in terms of different rate-limiting steps for the formation of the measured products.

The effect of three cationic surfactants bearing the same polar head group and different chain length (cetyltrimethyl ammonium bromide (CTAB); tetradecyltrimethylammonium bromide (TTAB); dodecyltrimethylammonium bromide (DTAB)) on the conformation and function of the sea anemone pore-forming toxins sticholysins I and II (St I and St II) was studied by fluorescence and circular dichroism spectroscopy and evaluation of hemolytic activity (HA). Preincubation of the toxins with the longer chain surfactants CTAB and TTAB at concentrations slightly above their critical micelle concentration (CMC) leads to an enhancement of their HA. Significant increases in the fluorescence intensity with a slightly red shift in lambda(max) were observed at concentrations close to the surfactants' CMC, suggesting changes in the environment of the tryptophan residues. The changes in the fluorescence intensity are more noticeable and take place at lower surfactant concentrations for St I, irrespective of the surfactant alkyl chain length, although the differences between St I and St II increase as the surfactant alkyl chain length increases. This is evinced not only by the higher fluorescence intensity values and the lower surfactant concentrations required to reach them, but also by the higher acrylamide-quenching constant values (Ksv) for St I. However, the surfactant's effects on the toxins' HA were not found to be directly related to the observed changes in fluorescence intensity, as well as near- and far-UV-CD spectra. In particular, the latter spectra indicate that changes in HA and in fluorescence behavior take place without noticeable modifications in St I and St II secondary and tertiary structures. The results suggest that the interaction with the surfactants induces only subtle conformational changes in the toxins that favor the formation of lytic competent structures.