Probing of lipase activity at air/water interface by sum-frequency generation spectroscopy

The infrared-visible sum-frequency generation (SFG) vibrational spectroscopy was used to probe enzymatic activity of Thermomyces lanuginosus lipase (TLL) at air/water interface. A monolayer of amphiphilic O-palmitoyl-2,3-dicyanohydroquinone (PDCHQ), containing target ester group and two CN groups serving as vibrational markers, was utilized as an enzyme substrate. SFG data revealed the detailed molecular scale structure and properties of the PDCHQ layer at the interface. In particular, we demonstrate that hydrophilic headgroup of PDCHQ is mainly in the form of an oxyanion, and the enzyme-induced cleavage of the ester bond could be spectroscopically monitored by the disappearance of the intense C tripple bond N resonance at 2224 cm(-1). The enzymatic nature of the ester bond cleavage was confirmed by the control experiments with deactivated S146A mutant variant of TLL. By comparing action of wild type (WT) TLL and its inactive S146A mutant, it was shown that two effects take place at the interface: disordering of the lipid monolayer due to the adsorption of enzyme and enzymatic cleavage of the ester bond. The concentration of enzyme as low as 10 nM could be easily sensed by the SFG spectroscopy. We present spectroscopic evidence that upon hydrolysis one of the products, 2,3-dicyanohydroquinone, leaves the surface, while the other, palmitic acid, remains at air/water interface in predominantly undissociated form with the mono-hydrogen-bonded carbonyl group. Strong amide I (1662 cm(-1)) and amide A (3320 cm(-1)) SFG signals from TLL suggest that enzyme molecules position themselves at air/water interface in an orderly fashion. Presented work demonstrates the potential of SFG spectroscopy for in situ real-time monitoring of enzymatic processes at air/water interface.     

Anion configuration at the air/liquid interface of ionic liquid [bmim]OTf studied by sum-frequency generation spectroscopy

The air/liquid interface of a room temperature ionic liquid, 1-butyl-3-methylimidazolium trifluoromethanesulfonate ([bmim]OTf), is investigated using infrared-visible sum frequency generation (SFG) spectroscopy. The SFG spectra clearly show low-frequency modes [CF 3-symmetric stretching (ss) mode and SO 3-symmetric stretching (ss) mode] of the OTf anion, demonstrating the existence of anions polar oriented at the interface. The amplitude of the CF 3-ss peak of the OTf anion has the opposite sign with respect to that of the SO 3-ss peak, indicating that OTf anions at the surface have polar ordering where the nonpolar CF 3 group points away from the bulk into the air, whereas the SO 3 group points toward the bulk liquid. The line width of the SFG peak from the submerged SO 3 group is appreciably narrower than that from IR absorption, suggesting the environment of the surface OTf anions is much more homogeneous than that of the bulk. The vibrational calculations also suggest that the anions and the cations form a more specific aggregated configuration at the surface as compared to the bulk.     

Theory of time-resolved sum-frequency generation and its applications to vibrational dynamics of water

A molecular theory of time-resolved sum-frequency generation (SFG) has been developed. The theoretical framework is constructed using the coupled-oscillator model in the adiabatic approximation. This theory can treat not only the vibrational spectroscopy but also vibrational dynamics. An application of this theory is also provided for estimation of the time constants of the intermolecular vibrational energy transfer between water molecules. This approach can be used for molecular analysis of the experimental results of Shen at al. on the SFG studies of vibrational dynamics of water.     

Mixtures of sodium dodecyl sulfate/dodecanol at the air/water interface by computer simulations

Molecular dynamics simulations of monolayers of surfactant mixtures at the air/water interface were performed where the binary mixture was composed of sodium dodecyl sulfate (SDS) and dodecanol molecules. At the same ratio of SDS and dodecanol molecules, two monolayer mixtures were prepared. In the first monolayer, all the dodecanol molecules were placed together in the center of the simulation box, whereas in the second monolayer, those molecules were uniformly distributed in the surface area in such a way that they were far from each other. Simulations of both systems indicate that the dodecanol tails in the first monolayer are straighter and more ordered than those in the second monolayer. From the present results, we observed new insights of how the different molecules should array or distribute at the interface in real systems. Finally, studies of the interfacial water around the different surfactants were also analyzed, showing that they are closer to the polar headgroups of dodecanol than to the SDS headgroups.     

Structure and dynamics of water at water|Pt interface as seen by molecular dynamics computer simulation

Five molecular dynamics computer simulations were performed to study the structural and dynamical properties of water next to uncharged and charged Pt surfaces. The results show that the structure of a water layer adsorbed on the metal surface is very sensitive to the details of the water–metal potential. While patches of short-living hexagonal ice-like structure are observed in the adsorbed water layer next to the uncharged Pt(111) surface, a square lattice solid-like structure is seen for the layer on top of the uncharged Pt(100) surface. The orientational ordering for the following two layers of water next to uncharged Pt is displaying a preference towards the orientations that are characteristic of hexagonal ice-I, while water is liquid-like in these layers. In the presence of a high value external electric field water reorients and undergoes a layering transition.     

Surfactant layers at the air/water interface: structure and composition

The use of neutron reflectometry to study the structure and composition of surfactant layers adsorbed at the air/water interface is reviewed. A critical assessment of the results from this new technique is made by comparing them with the information available from all other techniques capable of investigating this interface.     

Study of the dynamics of surface molecules by time-resolved sum-frequency generation spectroscopy

Sum-frequency generation (SFG) is a nonlinear laser-spectroscopy technique suitable for analysis of adsorbed molecules. The sub-monolayer sensitivity of SFG spectroscopy enables vibrational spectra to be obtained with high specificity for a variety of molecules on a range of surfaces, including metals, oxides, and semiconductors. The use of ultra-short laser pulses on time-scales of picoseconds also makes time-resolved measurements possible; this can reveal ultrafast transient changes in molecular arrangements. This article reviews recent time-resolved SFG spectroscopy studies revealing site-hopping of adsorbed CO on metal surfaces and the dynamics of energy relaxation at water/metal interfaces. Time-resolved sum frequency generation spectroscopy at surfaces with non-resonant laser pulse irradiation     

Second-harmonic generation optical activity of a polypeptide alpha-helix at the air/water interface

Quantitative measurements of second-harmonic generation optical activity (SHG-OA) have been performed for alpha-helical polypeptides poly-(gamma-benzyl-L-glutamate) and poly-(gamma-ethyl-L-glutamate) adsorbed at the airwater interface, with the fundamental frequency variant Planck's over 2piomega = 2.96 eV (lambda = 417 nm). The chiral component of the nonlinear susceptibility chi(XYZ) ((2)) is small for both polymers, being comparable in magnitude with the susceptibility chi(XXZ) ((2)) of the clean airwater interface. The microscopic origin of the nonlinear response has been investigated by using semiempirical ZINDOS calculations in conjunction with standard time-dependent perturbation theory to evaluate the molecular hyperpolarizability tensor of a model alpha-helix composed of glycine residues. Calculated nonlinear susceptibilities (per monomer unit) are in good agreement with experimental measurements for both the chiral and achiral response. The computational results indicate that charge transfer transitions of the alpha-helix have a large influence on the achiral components of the hyperpolarizability tensor, and produce characteristic features in the response under suitable experimental conditions. The dominant origin of SHG-OA for the model alpha-helix is a structural effect due to the tilt of the plane of each amide group of the helix relative to the helical axis. SHG-OA is associated with the orientational distribution of isolated, achiral chromophores, and is present in the absence of electronic coupling between the amide subunits of the polypeptide alpha-helix.     

Molecular dynamics simulations of amphiphilic graft copolymer molecules at a water/air interface

Fully atomistic molecular dynamics simulations of amphiphilic graft copolymer molecules have been performed at a range of surface concentrations at a water/air interface. These simulations are compared to experimental results from a corresponding system over a similar range of surface concentrations. Neutron reflectivity data calculated from the simulation trajectories agrees well with experimentally acquired profiles. In particular, excellent agreement in neutron reflectivity is found for lower surface concentration simulations. A simulation of a poly(ethylene oxide) (PEO) chain in aqueous solution has also been performed. This simulation allows the conformational behavior of the free PEO chain and those tethered to the interface in the previous simulations to be compared.     

Ions at the air/water interface

In a recent review of this topic [B.C. Garett, Science 303 (2004) 1146] the emphasis was on some recent experiments, in which it was found that some anions accumulate at the air/water interface and not in the bulk, as usually happens to the cations, and on some simulations which explained those positive surface adsorption excesses. Because a large number of these experiments could be explained on the basis of some simple physical models proposed by the authors for the interaction between the ions and the air/water interface [M. Manciu, E. Ruckenstein, Adv. Colloid Interface Sci. 105 (2003) 63; Adv. Colloid Interface Sci. 112 (2004) 109; Langmuir 21 (2005) 11312], those models are reviewed in the present note, the goal being to draw attention to them.     

Chain conformation of poly(dimethyl siloxane) at the air/water interface by sum frequency generation

This is to report a study of chain conformation of poly(dimethylsiloxane) (PDMS) in spread monolayers at the air/water interface (A/W) with the aid of vibrational sum frequency spectroscopy (VSFS). We find that methyl groups of PDMS chains at the interface are completely disordered in the dilute regime of the surface density. At higher surface densities, however, the two methyl groups on the repeating unit point into the air asymmetrically; one points more normal to the interface, whereas the other lies more parallel to the interface. In the first collapsed regime, where the surface pressure of the PDMS monolayer reaches a plateau value of 8.7 mN/m, the signal intensity at 2915 cm (-1), assigned to the symmetric vibrational frequency of the methyl groups, is found independent of the surface density. On the basis of this finding, we propose that PDMS chains, in the first collapse regime at the A/W, form asymmetric layers. Thus, our proposal lends support to earlier works by Langevin's group to refute a widely speculated helix model that was based on energy minimization in the crystalline state of PDMS. In short, the energy consideration in the bulk crystalline state does not provide meaningful guidance as to the chain conformation of the monolayer at the A/W.     

Liquid/air interfacial structure of alcohol-octyl hydroxamic acid mixtures: a study by sum-frequency spectroscopy

The molecular structure of the liquid/air interfaces of 1-octanol, 1-decanol, n-decane and the branched decyl alcohol EXXAL 10 has been studied by sum-frequency spectroscopy (SFS) in the C-H stretching vibrational region. The data suggest that the interfaces consist of ordered molecules with closely packed alkyl tails, in close to all-trans conformation with some gauche defects. The degree of surface ordering for the branched alcohol is much higher than for octanol and decanol. When octyl hydroxamic acid (OHA) is dissolved in 1-octanol it increases the gauche conformational defects in the interfacial chains, possibly due to mixing with the surface alcohol molecules and disrupting their ordering. In contrast, we suggest that when octyl hydroxamic acid is dissolved in EXXAL 10, the surface ordering of the alcohol chains does not change. We put forward the hypothesis that the appearance of new bands, belonging to the asymmetric methylene group vibrations and to the asymmetric methyl modes in the SF spectra of the mixture suggests that the surface OHA molecules are arranged with their hydrocarbon tails tilted very close to the interface.     

Studies on the electrical double layer structure at the water/air interface

Effective dipole moments (calculated from experimental data of surface tension and electric surface potential) of some homologous normal alcohols and carboxylic acid were found to vary linearly with the number of carbon atoms in the hydrocarbon chain. Values of effective dipole moments were used for the determination of the effective dipole moments of water molecules , and the dielectric permittivity of the water subphase (₁), as well as in the vicinity of the hydrophobic part of adsorbed molecule (₂). The latter was found to decrease with the increase of the hydrocarbon chain length. Knowing the effective dipole moment of surface water dipoles, the average orientation angle () of water molecules at the inteface was estimated. The calculated potential drop of water varies within the range –0.038 to –2.38 V for two extreme orientations of water dipoles at the surface.     

Polarization and experimental configuration analyses of sum frequency generation vibrational spectra, structure, and orientational motion of the air/water interface

Here we report a detailed study on spectroscopy, structure, and orientational distribution, as well as orientational motion, of water molecules at the air/water interface, investigated with sum frequency generation vibrational spectroscopy (SFG-VS). Quantitative polarization and experimental configuration analyses of the SFG data in different polarizations with four sets of experimental configurations can shed new light on our present understanding of the air/water interface. Firstly, we concluded that the orientational motion of the interfacial water molecules can only be in a limited angular range, instead of rapidly varying over a broad angular range in the vibrational relaxation time as suggested previously. Secondly, because different vibrational modes of different molecular species at the interface has different symmetry properties, polarization and symmetry analyses of the SFG-VS spectral features can help the assignment of the SFG-VS spectra peaks to different interfacial species. These analyses concluded that the narrow 3693 cm(-1) and broad 3550 cm(-1) peaks belong to C(infinityv) symmetry, while the broad 3250 and 3450 cm(-1) peaks belong to the symmetric stretching modes with C2v symmetry. Thus, the 3693 cm(-1) peak is assigned to the free OH, the 3550 cm(-1) peak is assigned to the singly hydrogen-bonded OH stretching mode, and the 3250 and 3450 cm(-1) peaks are assigned to interfacial water molecules as two hydrogen donors for hydrogen bonding (with C2v symmetry), respectively. Thirdly, analysis of the SFG-VS spectra concluded that the singly hydrogen-bonded water molecules at the air/water interface have their dipole vector directed almost parallel to the interface and is with a very narrow orientational distribution. The doubly hydrogen-bonded donor water molecules have their dipole vector pointing away from the liquid phase.     

Orientational motions of vibrational chromophores in molecules at the air/water interface with time-resolved sum frequency generation

The first time-resolved experiments in which interfacial molecules are pumped to excited electronic states and probed by vibrational sum frequency generation (SFG) are reported. This method was used to measure the out-of-plane rotation dynamics, i.e. time dependent changes in the polar angle, of a vibrational chromophore of an interfacial molecule. The chromophore is the carbonyl group, the rotation observed is that of the -C=O bond axis, with respect to the interfacial normal, and the interfacial molecule is coumarin 314 (C314) at the air/water interface. The orientational relaxation time was found to be 220+/-20 ps, which is much faster than the orientational relaxation time of the permanent dipole moment axis of C314 at the same interface, as obtained from pump-second harmonic probe experiments. Possible effects on the rotation of the -C=O bond axis due to the carbonyl group hydrogen bonding with interfacial water are discussed. From the measured equilibrium orientation of the permanent dipole moment axis and the carbonyl axis, and knowledge of their relative orientation in the molecule, the absolute orientation of C314 at the air/water interface is obtained.     

Microscopic collective dynamics of water

Data obtained on microscopic collective excitations in water by molecular dynamics simulation within the framework of the coarse-grained mW-model of the intermolecular interaction potential for water are reported. The calculated spectra of the dynamic structure factor and spectral densities of time correlation functions of longitudinal and transverse currents reveal the existence of propagating collective excitations of longitudinal and transverse polarization in water for a wide range of wavenumbers. The dynamics of fluctuations in the particle number density is analyzed within the framework of a microscopic theory that takes into account only the structural features of a system. The theoretically calculated data on the spectra of dynamic structure factor in a wavenumber range of 0.13–0.48 Å^–1 are in good agreement with the results of molecular dynamics simulation.     

Spectroscopic investigation of Pt—methanol interface in perchloric acid medium by sum-frequency generation

We recorded vibrational spectra of the CO poison resulting from the electrochemical decomposition of CH₃OH on platinum by sum-frequency generation using the infrared free electron laser CLIO. At high CH₃OH concentration (0.1 M) in the electrolyte, both multi-bonded and linearly bonded CO are equally present. At lower CH₃OH concentration (0.004 M), the multi-bonded species is predominant. The CO poison still remains on the surface at potentials (0.05 V/NHE) close to the hydrogen evolution onset.     

On the interactions of ions with the air/water interface

In the vicinity of a charged interface, the Poisson-Boltzmann approach considers that the ions obey Boltzmann distributions in a mean electrical field that satisfies the Poisson equation. However, the boundary between two dielectrics generates additional interactions between ions and the interface. The traditional models of ion hydration interactions, that assume that water is a homogeneous dielectric, predict that these interactions are repulsive for all kinds of ions, since all ions should prefer the medium with a larger dielectric constant, where they are better hydrated. In reality, the interactions between the ions and the neighboring water molecules can generate additional short-range ion-hydration interactions, which are either repulsive (for structure-making ions) or attractive (for structure-breaking ions). In the present paper, various models for the ion-hydration forces are examined and compared with the results of molecular dynamics simulations. At large ionic strengths, the latter results could be reproduced qualitatively only when short-ranged attractions between the structure-breaking ions and the interface were taken into account.     

Interaction of nitrophenols with lipids at the air/water interface

The uptake of ortho and para nitrophenol to charged and neutral lipid monolayers spread at the air/solution interface was studied by reflection spectroscopy. The adsorption characteristics of the two nitrophenols have been studied by measuring the surface pressure and surface potential as a function of molecular area of the different lipid monolayers in the presence of nitrophenols in the subphase. The results have been interpreted in terms of the electrostatic interaction between the negatively charged dissociated phenolate ions and the positively charged head group of dioctadecyldimethylammonium bromide monolayers.