Reversed micellar system

In the past few years, a range of solvation dynamics experiments have been demonstrated for reverse micellar systems. Reverse micelles form when a polar solvent is sequestered by surfactant molecules in a continuous nonpolar solvent. The interaction of the surfactant polar headgroups with the polar solvent can result in the formation of a well-defined solvent pool. Many different kinds of surfactants have been used to form reverse micelles. However, the structure and dynamics of reverse micelles created with Aerosol-OT (AOT) have been most frequently studied. AOT reverse micelles are monodisperse, spherical water droplets [32]. The micellar size is directly related to the water volume-to-surfactant surface area ratio defined as the molar ratio of water to AOT,... 

The majority of practical micellar systems of Tionnal micelles use water as tire main solvent. Reverse micelles use water immiscible organic solvents, altlrough tire cores of reverse micelles are usually hydrated and may contain considerable quantities of water. Polar solvents such as glycerol, etlrylene glycol, fonnamide and hydrazine are now being used instead of water to support regular micelles [10]. Critical fluids such as critical carbon dioxide are... 

Fig. 2. CD-spectra of (L-Lys HBr) in reversed micellar systems of AOT/octane/H20 at different w0-values 65). T 20 °C...

As mentioned above, water structure in reversed micelles deviates considerably from the structure in the bulk-phase. Therefore, the hydration shell of macromolecules entrapped in reversed micellar systems should be changed and thus also their conformation. According to the results of several authors this is indeed the case. 

The conformation of bovine myelin basic protein (MBP) in AOT/isooctane/water reversed micellar systems was studied by Waks et al. 67). This MBP is an extrinsic water soluble protein which attains an extended conformation in aqueous solution 68 but is more density packed at the membrane surface. The solubilization of MBP in the AOT reversed micelles depends on the water/AOT-ratio w0 68). The maximum of solubilization was observed at a w0-value as low as 5.56. The same value was obtained for another major protein component of myelin, the Folch-Pi proteolipid 69). According to fluorescence emission spectra of MBP, accessibility of the single tryptophane residue seems to be decreased in AOT reversed micelles. From CD-spectra one can conclude that there is a higher conformational rigidity in reversed micelles and a more ordered aqueous environment. 

Investigations of the solubilization of water and aqueous NaCl solutions in mixed reverse micellar systems formed with AOT and nonionic surfactants in hydrocarbons emphasized the presence of a maximum solubilization capacity of water, occurring at a certain concentration of NaCl, which is significantly influenced by the solvent used [132],... 

In addition to solubilization, entrapment of polymers inside reversed micelles can be achieved by performing in situ suitable polymerization reactions. This methodology has some specific peculiarities, such as easy control of the polymerization degree and synthesis of a distinct variety of polymeric structures. The size and shape of polymers could be modulated by the appropriate selection of the reversed micellar system and of synthesis conditions [31,191]. This kind of control of polymerization could model and/or mimic some aspects of that occurring in biological systems. 

Size Exclusion Chromatography of Some Reversed Micellar Systems... 

Dagaonkar MV, Mehra A, Jain R, Heeres HJ (2004) Synthesis of CaC03 nanoparticles by carbonation Of lime solutions in reverse micellar systems. Chem Eng Res Des 82 (11) 1438-1443... 

The partial racemization of isolated 2-octanol suggests that the hydrolysis may proceed via ionization of optically active substrates as in the Sjjl hydrolysis in homogeneous solution. The hydrolysis via ionization may be suppressed in media with low dielectric constant like micelles (Okamoto and Kinoshita, 1972), resulting in net retention. The ineffectiveness of the stereochemical influence of the CTAB micelle may be interpreted as a consequence of the mutual repulsion of the positively charged head groups of [46] and CTAB, so there is need for molecules of solvent to be incorporated between surfactant head groups (Sukenik et al., 1975). An appreciable increase in retention was also observed in a reversed micellar system (Kinoshita and Okamoto, 1977). 

Recently however, it has been recognized that liquid extraction is a potential method in the primary recovery of fermentation cell culture products, such as proteins and amino acids. The separation problem, however, is difficult because the product mixtures are often complex, including cell debris and enzymes. Proteins are not suitable for conventional solvent extraction because of incompatibility with organic solvents, but can be handled in aqueous two-phase systems or by extraction in reverse micellar systems (Chapter 15). 

Reactivities of pentacyanoferrates(II) in micelles and reversed micelles have been studied. The hexadecyltrimethylammonium cation causes a modest increase in rate constant for the anion-anion reaction [Fe(CN)5(4-CNpy)] + CN. This can equally well be interpreted according to the pseudophase model developed from the Olson-Simonson treatment of kinetics in micellar systems or by the classical Bronsted equation. 

Rao AM, John VT, Gonzalez RD, Akkara JA, Kaplan DL. Catal3dic and interfacial aspects of enzymatic polymer synthesis in reversed micellar systems. Biotechnol Bioeng 1993 41 531-540. 

Jain TK, Cassin G, Badiali JP, PUeni MP (1996) Relation between exchange process and structure of ACT reverse micellar system. Langmuir 12 2408-2411... 

Osseoasare K, Arriagada FJ (1990) Preparation of Si02 Nanoparticles in a Nonionic Reverse Micellar System. Colloids Surf 50 321-339... 

Although RMs are thermodynamically stable, they are highly dynamic. The RMs constantly colhde with each other and occasionally a colhsion results in the fusion of two RMs temporarily. During this fusion surfactant molecules and the contents residing inside RMs may be exchanged. In AOT reverse micellar system, this dynamic behavior exhibits second-order kinetics with rate constants in the order of 10 to 10 M s [37]. This dynamic nature not only influences the properties of the bulk system but also affects the enzymatic reaction rates [38]. 

In order to be exploitable for extraction and purification of proteins/enzymes, RMs should exhibit two characteristic features. First, they should be capable of solubilizing proteins selectively. This protein uptake is referred to as forward extraction. Second, they should be able to release these proteins into aqueous phase so that a quantitative recovery of the purified protein can be obtained, which is referred to as back extraction. A schematic representation of protein solubilization in RMs from aqueous phase is shown in Fig. 2. In a number of recent publications, extraction and purification of proteins (both forward and back extraction) has been demonstrated using various reverse micellar systems [44,46-48]. In Table 2, exclusively various enzymes/proteins that are extracted using RMs as well as the stability and conformational studies of various enzymes in RMs are summarized. The studies revealed that the extraction process is generally controlled by various factors such as concentration and type of surfactant, pH and ionic strength of the aqueous phase, concentration and type of CO-surfactants, salts, charge of the protein, temperature, water content, size and shape of reverse micelles, etc. By manipulating these parameters selective sepa-... 

Table 2. Enzymes/proteins studied in reverse micellar systems... 

Biomolecule Source Reverse micellar system Nature of study Reference... 

---