Micelles water pools

Effect of salt type and concentration The ionic strength of the aqueous solution in eontaet with a reverse micelle phase affects protein partitioning in a number of ways [18,23]. The first is through modification of electrostatic interactions between the protein surface and the surfaetant head groups by modifieation of the eleetrieal double layers adjacent to both the eharged inner mieelle wall and the protein surface. The second effect is to salt out the protein from the mieelle phase because of the inereased propensity of the ionie speeies to migrate to the micelle water pool, reduee the size of the reverse mieelles, and thus displace the protein. 

For small amounts of solubilized water, as a polar additive, the stability of the micelle is markedly increased, as shown by a decrease in the CMC. On the other hand, large amounts of water as a polar additive decrease the stability of the micelle. It is known that a solution of AOT in iso-octane solubilized up to 50 moles of water per mole of surfactant. As the concentration of water increases, the isotropic reverse micellar solution changes to a water-in-oil microemulsion. A clear understanding of the complex analyte-micelle-water pool interactions, especially analyte concentration and pH at the head group interfacial region, is under intensive study (Cline Love and al., 1984 Little and Singleterry, 1964 Luisi and Straub, 1984 Mclntire, 1990). 

In two-phase systems, however, where surfactant and water can partition between a fluid and a liquid phase, significant pressure effects occur. These effects were studied for AOT in ethane and propane by means of the absorption probe pyridine N-oxide and a fluorescence probe, ANS (8-anilino-l-naphthalenesulfonic acid) [20]. The UV absorbance of pyridine A-oxide is related to the interior polarity of reverse micelles, whereas the fluorescence behavior of ANS is an indicator of the freedom of motion of water molecules within reverse micelle water pools. In contrast to the blue-shift behavior of pyridine N-oxide, the emission maximum of ANS increases ( red shift ) as polarity and water motion around the molecule increase. At low pressures the interior polarity, degree of water motion, and absorbance intensity are all low for AOT reverse micelles in the fluid phase because only small amounts of surfactant and water are in solution. As pressure increases, polarity, intensity, and water motion all increase rapidly as large amounts of surfactant and water partition to the fluid phase. The data indicate that the surfactant partitions ahead of the water thus there is a constant increase in size and fluidity of the reverse micelle water pools up to the one-phase point. An example of such behavior is shown in Fig. 4 for AOT in propane with a total fVo of 40. The change in the ANS emission maximum suggests a continuous increase in water mobility, which is due to increasing fVo in the propane phase, up to the one-phase point at 200 bar. 

C. Physical Properties of Water in AOT Reverse Micelle Water Pools... 

Studies of water-in-scC02 reverse micelles derived from the surfactant ammonium carboxylate perfluoropolyether (PFPE) demonstrated that CO2 can partition into the PFPE reverse micelle [21], The important question related to the reaction rates and outcomes inside the nanoreactor is whether the pH within the reverse micelle water pool can be controlled by adjusting the CO2 pressure. In an attempt to answer this qnestion. Bright et al. quantitatively determined the pH within the PFPE water pools. Their results showed that the pH remains essentially constant as a fnnction of CO2 continnons phase pressure and micelle water loading [22]. 

Q. Zhong, A. P. Baronavski, and J. C. Owrutsky, J. Chem. Phys., 119,9171 (2003). Reorientation and Vibrational fhcrgy Relaxation of Pseudohahde Ions Confined in Reverse Micelle Water Pools. 

The strong interactions between the water molecules also become obvious from NMR measurements by Tsujii et al..57) 13C-NMR experiments were used for determining the microviscosity of water in reversed micelles of dodecylammonium-propionate with 13C glycine cosolubilized. It was found that the apparent viscosity of the water-pool corresponds to the viscosity of a 78 % aqueous glycerol solution, obviously as a consequence of the extended network formation by strong hydrogen bonding. 

The observation of slow, confined water motion in AOT reverse micelles is also supported by measured dielectric relaxation of the water pool. Using terahertz time-domain spectroscopy, the dielectric properties of water in the reverse micelles have been investigated by Mittleman et al. [36]. They found that both the time scale and amplitude of the relaxation was smaller than those of bulk water. They attributed these results to the reduction of long-range collective motion due to the confinement of the water in the nanometer-sized micelles. These results suggested that free water motion in the reverse micelles are not equivalent to bulk solvation dynamics. 

Investigation of water motion in AOT reverse micelles determining the solvent correlation function, C i), was first reported by Sarkar et al. [29]. They obtained time-resolved fluorescence measurements of C480 in an AOT reverse micellar solution with time resolution of > 50 ps and observed solvent relaxation rates with time constants ranging from 1.7 to 12 ns. They also attributed these dynamical changes to relaxation processes of water molecules in various environments of the water pool. In a similar study investigating the deuterium isotope effect on solvent motion in AOT reverse micelles. Das et al. [37] reported that the solvation dynamics of D2O is 1.5 times slower than H2O motion. 

The aqueous cores of reverse micelles are of particular interest because of their analogy with the water pockets in bioaggregates and the active sites of enzymes. Moreover, enzymes solubilized in reverse micelles can exhibit an enhanced catalytic efficiency. Figure B4.3.1 shows a reverse micelle of bis(2-ethylhexyl)sulfosuccinate (AOT) in heptane with three naphthalenic fluorescent probes whose excited-state pK values are much lower than the ground-state pK (see Table 4.4) 2-naphthol (NOH), sodium 2-naphthol sulfonate (NSOH), potassium 2-naphthol-6,8-disulfonate (NSOH). The spectra and the rate constants for deprotonation and back-recombination (determined by time-resolved experiments) provide information on the location of the probes and the corresponding ability of their microenvironment to accept a proton , (i) NDSOH is located around the center of the water pool, and at water contents w = [H20]/[A0T] >... 

Fig. B4.3.1. Schematic illustration of the average residence sites of the probes NOH (1), NSOH (2), NDSOH (3) in AOT reverse micelles. Length of the surfactant 11 A. Diameter of the water pool 18A at w = 3, 36A at w = 9. Largest dimension of the naphthol derivatives 9 A (adapted from Bardez et al.a ).

Figure 1 shows a reversed micelle where the bulk solvent is a hydrocarbon and the core is a water pool surrounded by surfactant. These systems possess unique features as the physical properties of the water pools only start to approach those of bulk water at high water content when the pool radii are >150 pools with radii as small as 15 can be constructed (1, 25). These systems have been used to investigate the nature of several inorganic reactions by stopped flow methods (26, 27). They have also been used to produce so-called naked ions, i.e., ions that possess a minimum of aqueous solvation (28). The system strongly promotes many reactions, a fact attributed to the unusual nature of the water in this system. 

Usually, activities of enzymes (hydrogenases included) are investigated in solutions with water as the solvent. However, enhancement of enzyme activity is sometimes described for non-aqueous or water-limiting surroundings, particular for hydrophobic (or oily) substrates. Ternary phase systems such as water-in-oil microemulsions are useful tools for investigations in this field. Microemulsions are prepared by dispersion of small amounts of water and surfactant in organic solvents. In these systems, small droplets of water (l-50nm in diameter) are surrounded by a monolayer of surfactant molecules (Fig. 9.15). The water pool inside the so-called reverse micelle represents a combination of properties of aqueous and non-aqueous environments. Enzymes entrapped inside reverse micelles depend in their catalytic activity on the size of the micelle, i.e. the water content of the system (at constant surfactant concentrations). 

Reverse micelles of CTAB in octane with hexanol as cosurfactant were reported to be able to lyse whole cells quickly and accommodate the liberated enzyme rapidly into the water pool of surfactant aggregates [50,51]. In another case a periplasmic enzyme, cytochrome c553, was extracted from the periplasmic fraction using reverse micelles [52]. The purity achieved in one separation step was very close to that achieved with extensive column chromatography. These results show that reverse micelles can be used for the extraction of intracellular proteins. 

Surfactants can aggregate in nonpolar solvents in the presence of small amounts of water with the tails oriented towards the bulk nonpolar solution and head groups interacting with water in the center (Fig. 2). The water pool formed in reverse micelles has been used as a medium to study chemical and biological reactions [22]. 

Photoinduced electron transfer from eosin and ethyl eosin to Fe(CN)g in AOT/heptane-RMs was studied and the Hfe time of the redox products in reverse micellar system was found to increase by about 300-fold compared to conventional photosystem [335]. The authors have presented a kinetic model for overall photochemical process. Kang et al. [336] reported photoinduced electron transfer from (alkoxyphenyl) triphenylporphyrines to water pool in RMs. Sarkar et al. [337] demonstrated the intramolecular excited state proton transfer and dual luminescence behavior of 3-hydroxyflavone in RMs. In combination with chemiluminescence, RMs were employed to determine gold in aqueous solutions of industrial samples containing silver alloy [338, 339]. Xie et al. [340] studied the a-naphthyl acetic acid sensitized room temperature phosphorescence of biacetyl in AOT-RMs. The intensity of phosphorescence was observed to be about 13 times higher than that seen in aqueous SDS micelles. 

Reverse micelles are small (1-2 nm in diameter), spherical surfactant aggregates huilt in an apolar solvent (usually referred to as oil), whereby the polar heads form a polar core that can contain water - the so-called water pool. The connection with autopoiesis is historically important, because it was with the collaboration with Francisco Varela that the work started (in fact it began as a theoretical paper - see Luisi and Varela, 1990). The idea was this to induce a forced micro-compartmentalization of two reagents, A and B, which could react inside the boundary (and not outside) to yield as a product the very surfactant that builds the boundary (Figure 7.13). The product S would concentrate at the membrane interface, which increases its size. Since reverse micelles are usually thermodynamically stable in only one given dimension, this increase of the size-to-volume ratio would lead to more micelles. Thus the growth and multiplication would take place from within the structure of the spherically closed unit, be governed by the component production of the micellar structure itself, and therefore (as will be seen better in... 

Figure 7.13 The first self-reproduction scheme conceived for reverse micelles, (a) A reverse micelle, (b) Two reagents, A and B, penetrate inside the water pool and react with each other inside the boundary, yielding the very surfactant S that makes the micelles. The S thus produced migrates to the boundary and induces growth and eventually multiplication of the micelle. (Adapted from Luisi and Varela, 1990).

Reverse micelles form in aprotic organic solvents, such as hydrocarbons or CCI4, and can be seen as a core containing water (the water pool) solubilized in an oily environment (for example hydrocarbons) by the hydrophobic tails. Figure 9.9 also shows the structure of AOT (from aerosol octyl), which is the most popular surfactant for reverse micelles. A typical reverse micellar system appears as a clear... 

Figure 9.10 Some structural details and dynamic properties of reverse micelles 50 irtM AOT/isooctane, Wo = 11.1 (= 10 p lHoOperml), 25°C 3.2% AOT (w/w), 1.4% H2O (w/w) mean water pool radius 20 A, mean hydrohynamic radius 32 A concentration of micelles 400 (xM, monomer AOT concentration 0.6-0.9 mM aggregation number 125 total interfacial area 14 m mC (Adapted from Fletcher and Robinson, 1981, and Harada and Schelly, 1982.)...

Figure 9.11 A case of selective compartmentation in reverse micelles, permitting the synthesis of a peptide by the reverse protease action. The product C, produced in the water pool, is expelled into the outside hydrocarbon environment due to its insolubility in water. (Adapted from Barbaric and Luisi, 1981.)...

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