Cationic Reverse Micelles Research Articles

Mimicking cellular fusion in a microfluidic channel via time-resolved chemiluminescence

Cell–cell fusion is a highly regulated and dramatic cellular event which is key to cell development. Reverse micelles (RMs) consist of nanoscopic droplets of water delimited by a dynamic, but well-defined surfactant layer, which confers them the status of simplified mimics of cells. Here, we have used anionic and cationic reverse micelles as mimetic environments of a cell, and monitored their fusion in a microfluidic channel via time-resolved evolution of chemiluminescence resulting from the fusion of reverse micelles. As the intracellular environment is highly crowded compared to relatively dilute in vitro conditions, we have also monitored time-resolved chemiluminescence resulting from the fusion of reverse micelles containing a model crowding agent PEG1000 in their aqueous core. Inter-micellar attractive interactions and surfactant film flexibility around the aqueous core of the reverse micelles dictate the kinetic pathways of their fusion.

PHOTOPHYSICAL ASPECTS OF POLARITY SENSITIVE FLUOROPHORE IN CATIONIC REVERSE MICELLES

In recent years, the field of reverse micelles has witnessed a significant growth of interest, partly due to the finding that proteins, other biopolymers, and even bacterial cells can be solubilized in the reverse micellar systems. Among surfactants capable of forming water in oil microemulsion, the cationic surfactant cetyltrimethylammonium bromide (CTAB) has received particular attention because of its ability to solubilize relatively large amount of water in a variety of hydrophobic organic solvents. Due to versatile photochemical and photophysical properties of Ru(II) complexes, Ru(II)- phenanthroline and its derivatives were used in this present work to form CTAB cationic reverse micelles and studied to characterise their photophysical properties such as absorption, emission and excited state life time. The findings from the spectral data observed in the present study proves that the excited state properties of Ru(II)-phenanthroline complexes are dramatically altered in the presence of reverse micelle. Specifically reverse micelle encapsulation of [Ru(dpphen)3]2+ complex in restricted environment causes blue shift in the emission maximum as well as have longer radiative lifetime. The blue shift in the emission maximum in the case of [Ru(dpphen)3]2+ indicates that the probes are near the water in oil reverse micellar interface, tightly bound and are not displaced towards the water pool of the micelle even at the highest water loading W0=50, which reveals the increased hydrophobicity. This highest fluorescence lifetime can serve as an excellent indicator to point out the location of the probe in a microheterogeneous system/environment.

Modified reverse micelle method as facile way to obtain several gold nanoparticle morphologies

Herein, we report a simple modified methodology based on cationic reverse micelles to form gold nanoparticles (AuNPs) with different morphologies and surface plasmon resonance (SPR) according to the phototherapeutic window (650–850 nm), to be applied in nanomedicine. In this sense, with available and simple changes, it was possible to obtain triangles, rods, spheres, truncated-decahedrons, and rings of gold nanoparticles. These structures were obtained with clear crystallinity changes on the different faces in the same solution, with two well-defined and intense absorption bands, which are interesting and applicable in cancer treatments through photothermic therapy.

Modulation of Kinetic Pathways of Enzyme-Substrate Interaction in a Microfluidic Channel: Nanoscopic Water Dynamics as a Switch.

Enzyme-mediated catalysis is attributed to enzyme-substrate interactions, with models such as "induced fit" and "conformational selection" emphasizing the role of protein conformational transitions. The dynamic nature of the protein structure, thus, plays a crucial role in molecular recognition and substrate binding. As large-scale protein motions are coupled to water motions, hydration dynamics play a key role in protein dynamics, and hence, in enzyme catalysis. Here, microfluidic techniques and time-dependent fluorescence Stokes shift (TDFSS) measurements are employed to elucidate the role of nanoscopic water dynamics in the interaction of an enzyme, α-Chymotrypsin (CHT), with a substrate, Ala-Ala-Phe-7-amido-4-methylcoumarin (AMC) in the cationic reverse micelles of benzylhexadecyldimethylammonium chloride (BHDC/benzene) and anionic reverse micelles of sodium bis(2-ethylhexyl)sulfosuccinate (AOT/benzene). The kinetic pathways unraveled from the microfluidic setup are consistent with the "conformational selection" fit for the interaction of CHT with AMC in the cationic reverse micelles, whereas an "induced fit" mechanism is indicated for the anionic reverse micelles. In the cationic reverse micelles of BHDC, faster hydration dynamics (≈550 ps) aid the pathway of "conformational selection", whereas in the anionic reverse micelles of AOT, the significantly slower dynamics of hydration (≈1600 ps) facilitate an "induced fit" mechanism for the formation of the final enzyme-substrate complex. The role of water dynamics in dictating the mechanism of enzyme-substrate interaction becomes further manifest in the neutral reverse micelles of Brij-30 and Triton X-100. In the former, the faster water dynamics aid the "conformational selection" pathway, whereas the significantly slower dynamics of water molecules in the latter are conducive to the "induced fit" mechanism in the enzyme-substrate interaction. Thus, nanoscopic water dynamics act as a switch in modulating the pathway of recognition of an enzyme (CHT) by the substrate (AMC) in reverse micelles.

Catanionic Reverse Micelles as an Optimal Microenvironment To Alter the Water Electron Donor Capacity in a SN2 Reaction.

The effect of interfacial water entrapped in two types of catanionic reverse micelles (RMs) on the kinetic parameters of the SN2 reaction between dimethyl-4-nitrophenylsulfonium trifluoromethanesulfonate (S+) and n-butylamine (BuNH2) was explored. Two catanionic surfactants, composed of a mixture of oppositely charged ionic surfactants without their original counterions, were used to create the RMs. Thus, benzyl- n-hexadecyldimethylammonium 1,4-bis(2-ethylhexyl) sulfosuccinate (BHD-AOT) and cetyltrimethylammonium 1,4-bis(2-ethylhexyl) sulfosuccinate (CTA-AOT) were formed. Also, the well-known anionic surfactant sodium 1,4-bis(2-ethylhexyl) sulfosuccinate (Na-AOT) was employed as a comparison. Our results showed an important catalytic-like effect of all RMs investigated in comparison with a water-benzene mixture, and the rate constant values depend on the type of surfactant used. Faster reaction in BHD-AOT RMs than in CTA-AOT and Na-AOT RMs was observed. This behavior was attributed to the strong interaction (by hydrogen bonding with AOT anion and ion-dipole interaction with BHD+) between the entrapped water and the BHD-AOT interface, which reduces the solvation capacity of water on S+. In CTA-AOT (and Na-AOT) RMs, the water-interface interaction is weaker and the electron pairs of water can solvate S+ ions. In summary, the chemical structure of the counterion on the catanionic surfactant alters the interfacial region, allowing the progress of a reaction inside the RMs to be controlled.

Subtleties of catanionic surfactant reverse micelle assemblies revealed by a fluorescent molecular probe

In this work, the absorption and emission behavior of the cationic hemicyanine trans-4-[4-(dimethylamino)styryl]-N-methylpyridinium iodide (HC) in reverse micelles (RMs) formed by the catanionic surfactants benzyl-n-hexadecyldimethylammonium-1,4-bis-2-ethylhexylsulfosuccinate (AOT-BHD) and cetyltrimethylammonium-1,4-bis-2-ethylhexylsulfosuccinate (AOT-CTA) have been investigated. Our results show that the spectroscopic behavior of HC changes when the dye is dissolved in AOT-BHD or in AOT-CTA RMs. While HC undergoes an intramolecular charge-transfer process upon excitation in AOT-CTA RMs, in AOT-BHD RMs this process is inhibited due to a specific interaction between HC and the polar head group of the BHD+ cation. This implies that the chemical structure of CTA+ and BHD+ cations has a large impact on the excited stated from which HC emission occurs. Additionally, the structural difference between the two cations impacts on the water–RM interface interaction, which provides a way of controlling the solvation process in these RMs. Furthermore, differences in the interfacial fluidity between the two catanionic RMs is observed, a result that is particularly interesting with regard to these systems being used as nanoreactors.

Efficient separation and reuse of CI Reactive Blue 19 from dyeing effluent by solvent extraction

In order to reduce environmental pollution from dyeing effluent, the cationic reverse micelles were prepared and used for removal of anionic Color Index (CI) Reactive Blue 19. The effect of surfactants, dyes, temperature and ultrasonic treatment on dye removal was investigated, respectively. The conductivity of CI Reactive Blue 19 aqueous solution was measured during removal process. In addition, the photograph of CI Reactive Blue 19 was also obtained. Finally, the removed dyes were backward extracted and reused for dyeing cotton fabrics. The results indicated that the removal of dyes was increased with increasing surfactants concentration and temperature, while the dye concentration exhibited the reverse trend. The higher removal of dyes could be caused by using ultrasonic treatment. The conductivity of dye aqueous solution was decreased with increasing surfactant concentration for the removal of dyes. The CI Reactive Blue 19 from backward extraction was reused for dyeing cotton fabrics, which exhibited the good property.

Gold nanoparticles optical properties induced by water and an ionic liquid (bmimBF4) inside cationic reverse micelles

Polar solvent interactions at the interface are a key factor affecting the final properties of nanoparticles, including size, shape and crystallinity.

Effect of Cosurfactants on the Interfacial Hydration of CTAB Quaternary Reverse Micelle Probed Using Excited State Proton Transfer.

It has been proven previously that the negatively charged photoacid 8-hydroxypyrene-1,3,6-trisulfonic acid trisodium salt (HPTS) resides at the interface of the cationic reverse micelle (RM) cetyltrimethylammonium bromide (CTAB)/octanol/water/cyclohexane and is a potential reporter of hydration through the excited state proton transfer (ESPT) process. However, the ESPT dynamics monitored by the pump-probe study was limited to the ultrafast timescale and hence did not report any discernible ESPT signature. Herein, we reinvestigate the ESPT behavior using fluorescence spectroscopy in the nanosecond timescale and at different values of w0 (=[water]/[surfactant]). We clearly observed distinct w0-dependent ESPT signatures analogous to conventional ternary cationic RMs implying considerable interfacial hydration. The results agree with a recent molecular simulation study, where significant penetration of water molecules into the interface was predicted for the CTAB quaternary RM. Moreover, we also found that the ESPT dynamics and the fluorescence anisotropy decay of HPTS depend differentially on the octanol/CTAB ratio (p0). The ESPT process was found to be disfavored, whereas the anisotropy decay accelerates upon the increase in p0 values. Our analysis indicates that with the increase in the octanol concentration, dehydrated regions enrich gradually at the interface. However, the increase in octanol concentration may reduce the effective electrostatic potential experienced by the probe and thus may result in faster rotational relaxation.

Hydrophobically Tailored Carbon Dots toward Modulating Microstructure of Reverse Micelle and Amplification of Lipase Catalytic Response.

This article delineates the modulation of microstructure of cationic reverse micelle utilizing hydrophobically modified carbon dots (CDs) with varying surface functionalizations. Citric acid was used as the source of the carbon core, and Na-salt of glycine, glycine, Na-salt of 11-aminoundecanoic acid, 11-aminoundecanoic acid, and n-hexadecylamine were used for the surface fabrication of CDs to produce CD 1s, CD 1a, CD 2s, CD 2a, and CD 3, respectively. All these CDs having dimension of 5-7 nm were characterized by spectroscopic and microscopic techniques. The hydrodynamic diameter of cetyltrimethylammonium bromide (CTAB) reverse micelle (CTAB/isooctane/n-hexanol/water) at z ([cosurfactant]/[surfactant]) = 6.4 and W0 ([water]/[surfactant]) = 44 is around 15-20 nm. Interestingly, the size of the water-in-oil (w/o) microemulsions dramatically increased up to 120-200 nm upon doping hydrophobic surface functionalized CD 2a and CD 3. This is possibly due to change in the micellar exchange dynamics and reorganization of the micellar aggregates via hydrophobic interaction between surfactant (CTAB) tail and hydrophobic surface modifier of the carbon dots. However, no alteration in the size of reverse micelles was noted in the presence of carbon dots CD 1s, CD 1a, and CD 2s. Spectroscopic and microscopic investigations confirmed that the hydrophobic CD 2a and CD 3 were localized at the interface of reverse micelles whereas CD 1s, CD 1a, and CD 2s were possibly located in the water pool (away from interface). The activity of Chromobacterium viscosum lipase encapsulated within CD 3 and CD 2a doped significantly large CTAB reverse micelles showed remarkable improvement (3.7-fold and 3.4-fold) in its catalytic response. However, hydrophilic carbon dots CD 1s and CD 2s as well as moderately hydrophobic CD 1a had no significant effect on the microstructure of reverse micelles as well as on the lipase activity.

A protic ionic liquid, when entrapped in cationic reverse micelles, can be used as a suitable solvent for a bimolecular nucleophilic substitution reaction.

In this work, we have explored how the confinement of the protic ionic liquid (IL) ethylammonium nitrate (EAN) inside toluene/benzyl-n-hexadecyldimethylammonium chloride (BHDC) reverse micelles (RMs) affects the Cl(-) nucleophilicity on the bimolecular nucleophilic substitution (SN2) reaction between this anion and dimethyl-4-nitrophenylsulfonium trifluoromethanesulfonate. To the best of our knowledge this is the first report where toluene/BHDC RMs use EAN as a polar component and it is used as a nanoreactor for carrying out kinetic experiments. Dynamic light scattering results reveal the formation of RMs containing the protic IL. The kinetic results show that upon confinement, EAN becomes a suitable solvent for the SN2 reaction while in homogeneous media it is a bad option. Entrapped in BHDC RMs, due to the strong hydrogen bond interactions, EAN behaves as an aprotic-like IL which cannot deactivate the nucleophilic power of Cl(-) and yet increases the substrate solubility. These facts show the versatility of this kind of organized system to alter the polar solvent entrapped and its influence on the reaction rate when it is used as a nanoreactor.

Wet Interface of Benzylhexadecyldimethylammonium Chloride Reverse Micelle Revealed by Excited State Proton Transfer of a Localized Probe

Excited state proton transfer (ESPT) of an anionic photoacid 8-hydroxypyrene-1,3,6-trisulfonic acid (HPTS or pyranine) has been studied inside a cationic reverse micelle (RM), water/benzylhexadecyldimethylammonium chloride (BHDC)/benzene, using steady-state and time-resolved fluorescence spectroscopy. The observed ESPT behavior is found to be remarkably different from the known ESPT trend of HPTS inside anionic AOT and cationic CTAB RMs; the ESPT dynamics approaches that of bulk water at higher w0 (≥10) inside AOT RM while no ESPT was observed for CTAB reverse micelle [ Sedgwick J. Am. Chem. Soc. 2012 , 134 , 11904 - 11907 ]. The ESPT dynamics inside BHDC RM is remarkably slower compared to that of water at all w0 (= [water]/[surfactant]) values and relatively much less sensitive to w0 variation compared to AOT RM. 2D NOESY and fluorescence anisotropy measurements reveal that the probe (HPTS) is embedded inside the positive interface of BHDC RM. Despite its trapped location, HPTS is able to undergo ESPT due to significant penetration of water molecules into the interface. Furthermore, facile ESPT at higher w0 is consistent with higher degree of interface hydration as predicted by a recent MD simulation [ Agazzi Langmuir 2014 , 30 , 9643 - 9653 ]. The study shows that ESPT dynamics inside RM varies not only with the interface charge but also on the nature of the headgroup and solvation.

Molecular dynamics simulation of water/BHDC cationic reverse micelles. structural characterization, dynamical properties, and influence of solvent on intermicellar interactions.

We report results obtained from molecular dynamics (MD) experiments of benzylhexadecyldimethylammonium chloride (BHDC) cationic reverse micelles (RMs). In particular we analyzed equilibrium and dynamical characteristics of water/BHDC RMs in pure benzene, at two different water/BHDC ratios (W0 = 5 and W0 = 10). The RMs appear as elliptical aggregates with eccentricities close to ∼0.9. Analysis of the different spatial correlations reveals three different spatial domains in the RMs: a water inner pool, the surfactant interface, and the external solvent. The calculated accessible surface areas for the aqueous inner cores suggest a strong penetration of solvent molecules within the micellar interface domains. Comparison between the density profiles of both RMs shows an increment of the broadness in the distributions of all species at the interface, along with an increasing overlap between the tail segments of the surfactant and benzene molecules as one considers larger micelles. For the dynamical side, the rotational characteristic time scale for the confined water was found to be 1 order of magnitude larger than that of the bulk water. A similar effect was also observed for hydrogen bond dynamics. Both retardation effects diminish with the size of the aggregate. To the estimate the influence of the external solvent on the intermicellar interactions, free energy profiles for the coalescence process between RMs of similar size in pure benzene and in a n-heptane/benzene mixture were also investigated. The results indicate that the association process is facilitated by the presence of n-heptane in the external nonpolar phase. Comparison with previous theoretical and experimental results is also carried out.

Effect of the cationic surfactant moiety on the structure of water entrapped in two catanionic reverse micelles created from ionic liquid-like surfactants.

The behavior of water entrapped in reverse micelles (RMs) formed by two catanionic ionic liquid-like surfactants, benzyl-n-hexadecyldimethylammonium 1,4-bis-2-ethylhexylsulfosuccinate (AOT-BHD) and cetyltrimethylammonium 1,4-bis-2-ethylhexylsulfosuccinate (AOT-CTA), was investigated by using dynamic (DLS) and static (SLS) light scattering, FTIR, and (1)H NMR spectroscopy techniques. To the best of our knowledge, this is the first report in which AOT-CTA has been used to create RMs and encapsulate water. DLS and SLS results revealed the formation of RMs in benzene and the interaction of water with the RM interface. From FTIR and (1)H NMR spectroscopy data, a difference in the magnitude of the water-catanionic surfactant interaction at the interface is observed. For the AOT-BHD RMs, a strong water-surfactant interaction can be invoked whereas for AOT-CTA this interaction seems to be weaker. Consequently, more water molecules interact with the interface in AOT-BHD RMs with a completely disrupted hydrogen-bond network, than in AOT-CTA RMs in which the water structure is partially preserved. We suggest that the benzyl group present in the BHD(+) moiety in AOT-BHD is responsible for the behavior of the catanionic interface in comparison with the interface created in AOT-CTA. These results show that a simple change in the cationic component in the catanionic surfactant promotes remarkable changes in the RMs interface with interesting consequences, in particular when using the systems as nanoreactors.

Water-in-oil microemulsion doped with gold nanoparticle decorated single walled carbon nanotube: Scaffold for enhancing lipase activity

The present work reports the development of water-in-oil (w/o) microemulsion doped with newly designed nanocomposite comprising of gold nanoparticle (GNP) decorated single walled carbon nanotube (SWNT). This nanocomposite included cationic reverse micelle was used to boost the catalytic activity of a surface-active enzyme, Chromobacterium viscosum lipase (CV lipase). SWNT was non-covalently dispersed using cetyltrimethylammonium bromide (CTAB), cetylalaninetrimethylammonium chloride (CATAC) while GNP was synthesized by reduction of HAuCl4 with reducing/stabilizing agent trisodium citrate. Counterion exchange between cationic SWNT dispersing agent and anionic capping agent of GNP led to the formation of GNP decorated SWNT (SWNT–GNP) nanocomposite. This newly developed SWNT–GNP included CTAB reverse micelle was characterized by several microscopic and spectroscopic techniques. Interfacially located SWNT–GNP included w/o microemulsion (confirmed from biphasic and fluorescence experiment) was used as a proficient host for enhancing the catalytic activity of lipase. Lipase activity within this self-assembled soft nanocomposite improved up to 3.9-fold (second order rate constant, k2=1694±16cm3g−1s−1) compared to standard CTAB reverse micelle (k2=433±7cm3g−1s−1). In case of cetyltripropyl ammonium bromide (CTPAB) based reverse micelle, the observed lipase activity improved to k2=2036±11cm3g−1s−1 in the presence of SWNT–GNP composite. Notably, this catalytic activity of lipase within SWNT–GNP included reverse micelle was till date the highest activity found in any w/o microemulsion. The attainment of flexibility in enzyme conformation at the augmented interface was verified using circular dichroism (CD) spectroscopy.

Reverse Micelle Mediated Bulk Liquid Membrane Separation of Platinum Gold and Silver From Real Samples

Simple and selective reverse micelle mediated bulk liquid membrane extraction procedure is developed for the separation and simultaneous pre-concentration of precious metals from valuable wastes like spent auto mobile catalytic converters and gold jewelry washings. Quantification has been carried out using inductively coupled plasma optical emission spectrometry (ICP-OES). Tri octyl methyl ammonium chloride (TOAC), a cationic hydrophobic surfactant, forms reverse micelles after dissolution in trichloroethylene facilitating the extraction of platinum, gold and silver ions into the bulk liquid membrane. The procedure is based on the formation of extractable hydrophobic ion-associates between TOAC cationic reverse micelles and anionic metal ions. These hydrophobic ion-associates which are transported into the bulk organic membrane from an aqueous feed phase are subsequently stripped into a receiving solution in a unitary process. Optimum conditions for the overall transport efficiency of precious metals have been evaluated. The accuracy of the procedure was validated by analyzing GBWAu-18 ore from faulted zone and PTM-1a nickel-copper sulphide matte certified reference materials. The procedure has been applied to spent catalytic converters and jewellery washings. Supplemental materials are available for this article. Go to the publisher's online edition of Separation Science & Technology to view the supplemental file.

Spectral features of 4,4-diaminodiphenyl sulfone in anionic and cationic inverted micelles

This Letter reports the photophysics of 4,4-diaminodiphenyl sulfone in anionic and cationic reverse micelles (RM). Decreased fluorescence intensity with significant blue shift of intramolecular charge transfer (ICT) fluorescence maximum and longer emission lifetime of the entrapped probe in motionally restricted water nano-pool of anionic RM could be observed. Bi-exponential decay in anionic RM appears due to polarity heterogeneity in the pool. In a nut shell, experiments establish that the probe is anchored in the micellar head group region of anionic RM vis-à-vis far away from the core in cationic RM. Increasing pool size show the opposite effect to the probe.

Electron donor ionic liquids entrapped in anionic and cationic reverse micelles. Effects of the interface on the ionic liquid–surfactant interactions

The behavior of two ionic liquids (ILs) with high electron donor ability such as 1-butyl-3-methylimidazolium trifluoromethanesulfonate (bmimTfO) and 1-butyl-3-methylimidazolium trifluoroacetate (bmimTfA) entrapped in anionic and cationic reverse micelles (RMs) was investigated using dynamic light scattering (DLS) and FT-IR spectroscopy. The systems studied were chlorobenzene/sodium 1,4-bis-2-ethylhexylsulfosuccinate (AOT)/bmimTfO, chlorobenzene/AOT/bmimTfA, chlorobenzene/benzyl-n-hexadecyldimethylammonium chloride (BHDC)/bmimTfO and chlorobenzene/BHDC/bmimTfA. DLS results reveal the formation of RMs containing bmimTfO and bmimTfA as polar components since the droplet size values increase as Ws (Ws = [IL]/[surfactant]) increases. To the best of our knowledge this is the first report where it is shown that both ILs are entrapped by AOT and BHDC surfactants to effectively create RMs. Furthermore, it is shown that the RMs consist of discrete spherical and non-interacting droplets of IL stabilized by the surfactants. The larger droplet size values and the larger changes obtained for bmimTfO entrapped in AOT and BHDC RMs in comparison with those for bmimTfA in both RMs can be explained considering the different IL-surfactant interactions. The FT-IR results suggest that the ionic interactions (with the surfactant polar head groups, surfactant counterions or with the IL counterions) are substantially modified upon confinement. These interactions produce segregation of ILs's ions altering the composition of the RM interfaces. These facts show the versatility of this kind of organized systems to alter the ionic organization, information that can be very important if these media are used as nanoreactors because unique microenvironments can be easily created simply changing the RM components and Ws.

Striking Improvement in Peroxidase Activity of Cytochrome c by Modulating Hydrophobicity of Surface‐Functionalized Gold Nanoparticles within Cationic Reverse Micelles

This work demonstrates a remarkable enhancement in the peroxidase activity of mitochondrial membrane protein cytochrome c (cyt c) by perturbing its tertiary structure in the presence of surface-functionalised gold nanoparticles (GNPs) within cetyltrimethylammonium bromide (CTAB) reverse micelles. The loss in the tertiary structure of cyt c exposes its heme moiety (which is buried inside in the native globular form), which provides greater substrate (pyrogallol and H(2)O(2)) accessibility to the reactive heme residue. The surfactant shell of the CTAB reverse micelle in the presence of co-surfactant (n-hexanol) exerted higher crowding effects on the interfacially bound cyt c than similar anionic systems. The congested interface led to protein unfolding, which resulted in a 56-fold higher peroxidase activity of cyt c than that in water. Further perturbation in the protein's structure was achieved by doping amphiphile-capped GNPs with varying hydrophobicities in the water pool of the reverse micelles. The hydrophobic moiety on the surface of the GNPs was directed towards the interfacial region, which induced major steric strain at the interface. Consequently, interaction of the protein with the hydrophobic domain of the amphiphile further disrupted its tertiary structure, which led to better opening up of the heme residue and, thereby, superior activity of the cyt c. The cyt c activity in the reverse micelles proportionately enhanced with an increase in the hydrophobicity of the GNP-capping amphiphiles. A rigid cholesterol moiety as the hydrophobic end group of the GNP strikingly improved the cyt c activity by up to 200-fold relative to that found in aqueous buffer. Fluorescence studies with both a tryptophan residue (Trp59) of the native protein and the sodium salt of fluorescein delineated the crucial role of the hydrophobicity of the GNP-capping amphiphiles in improving the peroxidase activity of cyt c by unfolding its tertiary structure within the reverse micelles.

The effect of different interfaces and confinement on the structure of the ionic liquid 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide entrapped in cationic and anionic reverse micelles

The behavior of the ionic liquid (IL) 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide ([bmim][Tf(2)N]) entrapped in two reverse micelles (RMs) formed in an aromatic solvent as dispersant pseudophase: [bmim][Tf(2)N]/benzyl-n-hexadecyldimethylammonium chloride (BHDC)/chlorobenzene and [bmim][Tf(2)N]/sodium 1,4-bis-2-ethylhexylsulfosuccinate (AOT)/chlorobenzene, was investigated using dynamic light scattering (DLS), FT-IR and (1)H NMR spectroscopies. DLS results reveal the formation of RMs containing [bmim][Tf(2)N] as a polar component since the droplet size values increase as the W(s) (W(s) = [[bmim][Tf(2)N]]/[surfactant]) increases. Furthermore, it shows that the RMs consist of discrete spherical and non-interacting droplets of [bmim][Tf(2)N] stabilized by the surfactants. Important differences in the structure of [bmim][Tf(2)N] entrapped inside BHDC RMs, in comparison with the neat IL, are observed from the FT-IR and (1)H NMR measurements. The electrostatic interactions between anions and cations from [bmim][Tf(2)N] and BHDC determine the solvent structure encapsulated inside the nano-droplets. It seems that the IL structure is disrupted due to the electrostatic interaction between the [Tf(2)N](-) and the cationic BHDC polar head (BHD(+)) giving a high ion pair degree between BHD(+) and [Tf(2)N](-) at a low IL content. On the other hand, for the AOT RMs there is no evidence of strong IL-surfactant interaction. The electrostatic interaction between the SO(3)(-) group and the Na(+) counterion in AOT seems to be stronger than the possible [bmim](+)-SO(3)(-) interaction at the interface. Thus, the structure of [bmim][Tf(2)N] encapsulated is not particularly disrupted by the anionic surfactant at all W(s) studied, in contrast to the BHDC RM results. Nevertheless, there is evidence of confinement in the AOT RMs because the [bmim](+)-[Tf(2)N](-) interaction is stronger than in bulk solution. Thus, the IL is more associated upon confinement. Our results reveal that the [bmim][Tf(2)N] structure can be modified in a different manner inside RMs by varying the kind of surfactant used to create the RMs and the IL content (W(s)). These facts can be very important if these media are used as nanoreactors because unique microenvironments can be easily created by simply changing the RM components and W(s).