Cetyltrimethylammonium Bromide Reverse Micelles Research Articles

CTAB Reverse Micelles as Catalysts for the Oxidation of Ascorbic Acid by K3[Fe(CN)6

The oxidation of ascorbic acid by K3[Fe(CN)6] was studied in reverse micellar systems composed of CTAB (Cetyltrimethylammonium bromide), and it was found the observed first order (k1(aq) = 5.2×10−5 s−1, k1(rev) = 61.4×10−4 s−1) rate constant in reverse micellar medium is around forty times higher compared to aqueous medium under identical conditions. The rate enhancement (k2(aq) = 0.9×10−5 mole−1.dm3.sec−1, k2(rev) = 1.75×10−3 mole−1.dm3.sec−1) is attributed to the large concentration effect and lower dielectric constant in the reverse micelles. The rate of the reaction increases with increase in W = {[H2O]/[surfactant]} which is explained in terms of ionic strength of the water pool. The effect of surfactant concentration on rate was explained on the basis of Berezin pseudo phase model. Copyright © 2021 by Authors, Published by BCREC Group. This is an open access article under the CC BY-SA License (https://creativecommons.org/licenses/by-sa/4.0).

Kinetic Study of SN2 Reaction between Paranitrophenyl Benzoate and Hydrazine in the Presence of CTAB Reverse Micelles

Kinetic study of the reaction between p-Nitrophenyl benzoate (PNPB) by hydrazine (HYN) in the presence of Cetyltrimethylammonium bromide (CTAB)/Chloroform/Hexane reverse micellar medium shows that the reaction obeys first order kinetics with respect to each of the reactants. The rate of the reaction is much slower in reverse micellar medium compared to aqueous medium under identical conditions (kˈAq = 2.84×10−3 sec−1, krm =1.34×10−4 sec−1). The rate constants for the reaction in the reverse micellar medium have been determined at different values of W {W=[H2O]/[CTAB]} and at different concentrations of CTAB. It was found that the observed rate constant decreases with W. This kinetic behaviour was interpreted by using modified Berezin pseudo phase model, taking into consideration the distribution of the reactants, PNPB and hydrazine between the three pseudo phases, i.e., water pool, interface an organic phase. Copyright © 2021 by Authors, Published by BCREC Group. This is an open access article under the CC BY-SA License (https://creativecommons.org/licenses/by-sa/4.0).

Preparation of γ-Al2O3 membranes for ultrafiltration by reverse micelles-mediated sol-gel process

Sol-gel process was modified by reverse micelles (RMs) for the preparation of γ-Al2O3 ultrafiltration (UF) membranes and their applications in the efficient rejection of methyl blue and bovine serum albumin were explored. The optimized boehmite sol with a narrower particle size distribution and better sphericity was synthesized by using cetyltrimethyl ammonium bromide RMs as microreactors and was dip-coated on α-Al2O3 microfiltration substrates to prepare γ-Al2O3 UF membranes. It was found that not only the sol particle size, it's distribution and morphology had also an important effect on the pore size, pore size distribution, porosity and pore volume of the membrane. Additionally, the as-prepared membranes with a MWCO of 8 kDa exhibited a narrow pore size distribution (4.8–6.8 nm), relatively high pure-water flux (30.4 L/(h m2 bar)) and excellent rejection for bovine serum albumin and methyl blue with the maximum rejection rate of 96.2% and 96.8% respectively.

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.

Effect of surfactant structure on reverse micellar extraction of ovalbumin

The reports about reverse micellar extraction of protein with hydrophobicity (such as ovalbumin) have been very limited; most of the studies are focused on extraction of protein with very limited hydrophobicity, such as bovine serum albumin (BSA). In the present study, we found that similar to BSA, ovalbumin could be transferred into CTAB (cetyltrimethylammonium bromide) and DTAB (dodecyltrimethylammonium bromide) reverse micelles most efficiently when pH reached 7 and recovered into water at pH 4.3. However, the transfer of ovalbumin molecules between water and organic phases was less efficient compared with BSA transfer. Noticing the surfactants used in reverse micellar extraction have been mostly conventional surfactants, in this study, we further used gemini surfactant pentamethylene-α,ω-bis(dimethyldodecylammonium bromide) reverse micelle to extract ovalbumin. We found the flexible spacer of the gemini surfactant could enhance ovalbumin transfer between the two phases and less amount of surfactant was needed. Besides, ovalbumin could be recovered from the gemini surfactant reverse micelle under wider and milder condition than BSA. Thus far the reports about the effects of surfactant structure and/or protein properties on protein extraction have been limited yet. This study may help us know more about how to optimize surfactant structure based on protein properties.

Dynamics of structural relaxation of stilbene 3 in solution, cetyltrimethylammonium bromide micelle, and bis(2-ethylhexyl) sulfosuccinate reverse micelle.

The relaxation dynamics of the excited states of stilbene 3 in various solvents, confined environment cetyltrimethylammonium bromide (CTAB) micelle, and water/bis(2-ethylhexyl) sulfosuccinate (AOT)/hexane reverse micelle are investigated. In the time-resolved decay curves of fluorescence measured in solution at excitation wavelength 256 or 266 nm, stilbene 3 underwent initially an ultrafast internal conversion to the S2 state or was directly excited at the S2 then via a conformational relaxation with time constant 1.1-4.6 ps. This relaxation process displays a linear dependence on solvent viscosity. Slow relaxation in deuterated methanol and water is explained that the vibrational energy is efficiently coupled to the librational modes of solvent. The conformational relaxation rate decreases slightly in the CTAB micelle but is greatly hindered in the AOT reverse micelle with small cavities. According to the results of anisotropy measurements, in both CTAB and AOT reverse micelles with a cavity diameter as large as ∼7 nm, the dynamics of molecular rotation remain significantly hindered.

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.

Graphene oxide in cetyltrimethylammonium bromide (CTAB) reverse micelle: A befitting soft nanocomposite for improving efficiency of surface-active enzymes

Herein, we report the successful inclusion of 2D allotrope of carbon, graphene oxide (GO) in cetyltrimethylammonium bromide (CTAB)/isooctane/n-hexanol/water reverse micelle without compromising the stability of water-in-oil (w/o) microemulsion. This newly developed self-assembled nanocomposites act as proficient host for surface-active enzymes, lipase, horseradish peroxidase (HRP), and soybean peroxidase (SBP). Lipase activity within GO-doped CTAB reverse micelles remarkably improved by 3.8-fold compared to that was observed in only CTAB reverse micelle (second-order rate constant, k2=433±7cm3g−1s−1). In case of GO-doped CTAB reverse micelle, the observed enzyme activity (k2=1653±11cm3g−1s−1) is till date the highest ever activity of lipase in CTAB w/o microemulsions. In case of HRP and SBP, the catalytic efficiency maximally increased up to 2.6-fold and 2.3-fold, respectively. Electrostatic attraction between cationic head group of CTAB and anionic surface of GO as well as intrinsic amphiphilic character of GO possibly resulted in the confinement of this 2D nanosheet at the interface of reverse micelles. Integration of GO at the interface augmented the interfacial space in vicinity of surface-active enzyme. This enlarged interface might have accommodated higher amount of substrate and lipase with flexibility in its conformation resulting in marked improvement in the enzyme activity. Interfacial localization of GO was established by fluorescence spectroscopy. In addition, change in secondary structure of lipase in presence of 2D carbon allotrope was substantiated by circular dichroism spectroscopy.

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.

Covalently Functionalized Single-Walled Carbon Nanotubes at Reverse Micellar Interface: A Strategy to Improve Lipase Activity

The present work reports covalent functionalization of single-walled carbon nanotubes (f-SWNTs) to introduce hydrophilicity to the otherwise amphiphobic nanotubes. The charge and spacer length of the functional moiety were varied by using quaternized ethylene diamine, 6-aminocaproate, quaternized (ethylenedioxy)bis(ethylamine), and a poly(ethylene glycol) (PEG) unit (f-SWNT-1 to f-SWNT-4, respectively). These f-SWNTs with varying degrees of hydrophilicity were incorporated within cetyltrimethyl ammonium bromide (CTAB) reverse micelles to develop stable self-assembled nanohybrids. An optimum hydrophilicity on the SWNT surface led to interfacial localization of f-SWNTs resulting in the augmentation of space at the interface. A surface-active enzyme, lipase, localized at this enhanced interface of f-SWNT-containing CTAB reverse micelles exhibited significant activation (2.5-fold) compared to that in the absence of the nanoconstructs. This improvement in lipase activity was mainly due to the smooth occupancy of lipase and also presumably because of the increase in the concentrations of both substrate and the enzyme at the augmented interface. Interestingly, the f-SWNTs that activate lipase in reverse micelles deactivate the same enzyme in water. The dispersion of f-SWNTs in water and its matching integration at the interface of reverse micelles were confirmed through transmission electron microscopic (TEM) investigations. The interfacial localization of these nanoconstructs was also established from the distinct fluorescence behavior of a hydrophobic fluorescent probe, fluorescein isothiocyanate (FITC), adsorbed onto the f-SWNT surface. In concurrence with the observed lipase activity, the corresponding changes in the enzyme conformation within f-SWNTs integrated reverse micelle as well as in aqueous medium were studied by circular dichroism (CD) and Fourier transform infrared (FTIR) spectroscopy.

GNP confinement at the interface of cationic reverse micelles: influence in improving the lipase activity

The present work reports thiol-assisted confinement of gold nanoparticles (GNPs) at the interface of reverse micelles with the aim to enhance the interfacial area and thereby the efficiency of surface-active Chromobacterium viscosum lipase. The strong gold capping ability of optimally hydrophobic thiols (1-dodecanethiol and 1,6-hexanedithiol) was aptly utilized to pull GNPs (∼3–5 nm) from the water pool to the oil/water interface of cetyltrimethylammonium bromide (CTAB) reverse micelles. These small sized GNPs were fitted at the microscopic interface of CTAB reverse micelles possibly because of the comparable thickness of the interface (∼1–2 nm) to that of the GNP diameter. Lipase solubilized within this augmented interface enjoys a flexible conformation, which resulted in the improvement of its activity (∼2.5 fold) with respect to only CTAB microemulsion. The activity of lipase within CTAB reverse micelles was thoroughly studied in the presence of mono and dithiols with varying chain length, where a greater improvement in activity was observed with dithiols. Bidentate ligand property of dithiols led to firm localization of higher number of GNPs at the interface which enhanced the total space in vicinity of enzyme at the interfacial domain. Fitting fusion of small sized GNPs within CTAB reverse micellar interface was confirmed by microscopic and spectroscopic studies. Smooth localization of lipase at the enhanced interface was also confirmed from the improvement in its secondary structure (α-helical content) in circular dichroism spectroscopic analysis. Interestingly, large sized GNPs (∼8 and 20 nm) were found to be well fitted at the interface of bigger head group-containing surfactants, cetyltriethylammonium bromide (CTEAB) and cetyltripropylammonium bromide (CTPAB). The hydrolytic efficiency of lipase in 1,6-hexanedithiol included GNP (∼20–25 nm)-doped CTPAB reverse micelles improved by ∼3.4 fold compared to that observed in only CTAB.

Enzyme immobilization on epoxy supports in reverse micellar media: Prevention of enzyme denaturation

Abstract Immobilization of enzymes such as α-chymotrypsin (EC 3.4.21.1), yeast alcohol dehydrogenase (YADH) from Saccharomyces cerevisiae (EC 1.1.1.1) and glucose dehydrogenase (GDH) from Gluconobacter cerinus (EC 1.1.1.119) has been carried out. Copolymers of allyl glycidyl ether (AGE) crosslinked with 25% ethylene glycol dimethacrylate (EGDM) (25 mg, dry wt) were contacted with the enzymes solubilized in reverse micellar media (0.5–5 mg/mL)overall of sodium bis(2-ethylhexyl) sulfosuccinate (AOT) salt in isooctane, and cetyl trimethylammonium bromide (CTAB) in chloroform–isooctane (50:50, v/v). Although the enzymes are readily denatured (>90%) after adsorption on the copolymer in aqueous buffers, no such adsorption-induced denaturation takes place in reverse micelles. α-Chymotrypsin is remarkably stable in AOT reverse micelles when 0.025 M citrate buffer of pH 9.0 containing 2 mM CaCl2 is used in the water pools instead of Tris–HCl buffer of pH 8.5. It was possible to achieve enzyme concentration of 5 mg/mL in 0.3 M AOT at molar ratio of water to surfactant, (W0), 30 and to obtain α-chymotrypsin loading of 20 mg/g of copolymer. The recovered enzyme solution can be reused with a fresh batch of polymer after supplementing the depleted solution. The immobilized enzyme exhibits excellent stability in aqueous buffers at room temperature and can be recycled several times. YADH is stable in both AOT and CTAB reverse micelles while GDH is stable only in CTAB reverse micelles containing 0.05 M Tris–HCl buffer of pH 8.5. Interestingly, the combination of YADH (2.5 mg/g) and GDH (0.5 mg/g) co-immobilized on the copolymer using CTAB–chloroform–isooctane system can be used for regeneration and recycle of NADPH at least 50 times as exemplified by complete reduction of a prochiral ketoester, ethyl 4-phenyl-2,4-dioxobutyrate (10 mM) to ethyl (R)-2-hydroxy-4-phenylbutyrate (HPB ester) using NADPH (0.2 mM).

Kinetics of oxidation of iodide by vanadium (V): Catalysis by the water pools of CTAB reverse micelles

The kinetic study of the oxidation of iodide ion by V(V) has been carried out in the water pools of cetyl trimethyl ammonium bromide(CTAB) reverse micelles in a mixture of chloroform-hexane (3 : 2). The study of the effect of concentration of V(V) and I− on rate show that the reaction obeys first order kinetics with each of the reactants. A plot of k′ (pseudo first order rate constant) versus [H+] is linear with a positive intercept at constant concentrations of iodide and bromide. The rate of the reaction is markedly increased in the reverse micellar medium compared to conventional aqueous medium under identical conditions. The pronounced acceleration in reverse micelles has been accounted for by the concentration effect in the water pool which have an effect on kinetics.

Influence of Gold Nanoparticles of Varying Size in Improving the Lipase Activity within Cationic Reverse Micelles

Herein, we report the effect of gold nanoparticles (GNPs) in enhancing lipase activity in reverse micelles of cetyltrimethylammonium bromide (CTAB)/water/isooctane/n-hexanol. The size and concentration of the nanoparticles were varied and their specific roles were assessed in detail. An overall enhancement of activity was observed in the GNP-doped CTAB reverse micelles. The improvement in activity becomes more prominent with increasing concentration and size of the GNPs (0-52 microM and ca. 3-30 nm, respectively). The observed highest lipase activity (k(2)=1070+/-12 cm(3) g(-1) s(-1)) in GNP-doped CTAB reverse micelles ([GNP]: 52 microm, ca. 20 nm) is 2.5-fold higher than in CTAB reverse micelles without GNPs. Improvement in the lipase activity is only specific to the GNP-doped reverse micellar media, whereas GNP deactivates and structurally deforms the enzyme in aqueous media. The reason for this activation is probably due to the formation of larger-sized reverse micelles in which the GNP acts as a polar core and the surfactants aggregate around the nanoparticle ('GNP pool') instead of only water. Lipase at the augmented interface of the GNP-doped reverse micelle showed improved activity because of enhancement in both the substrate and enzyme concentrations and increased flexibility in the lipase conformation. The extent of the activation is greater in the case of the larger-sized GNPs. A correlation has been established between the activity of lipase and its secondary structure by using circular dichroism and FTIR spectroscopic analysis. The generalized influence of GNP is verified in the reverse micelles of another surfactant, namely, cetyltripropylammonium bromide (CTPAB). TEM, dynamic light scattering (DLS), and UV/Vis spectroscopic analysis were utilized to characterize the GNPs and the organized aggregates. For the first time, CTAB-based reverse micelles have been found to be an excellent host for lipase simply by doping with appropriately sized GNPs.

A comprehensive study of extraction of hyperoside from Hypericum perforatum L. using CTAB reverse micelles

AbstractBACKGROUND: Hyperoside is a valuable natural pharmaceutical that has many biomedical functions. By the traditional method of alcohol extraction, only hyperoside solution of very low concentration can be obtained, so a new method is urgently needed to produce pure hyperoside more effectively. Reverse micellar extraction has been widely used in the purification of biological macromolecules. Theoretically, this method could also be used to purify materials with small molecules. Therefore it would seem appropriate to consider the extraction of hyperoside, a material with small molecules, using reverse micelles. In this study the factors affecting hyperoside extraction using cetyl trimethylammonium bromide (CTAB) reverse micelles were comprehensively investigated.RESULTS: The results showed that the partition coefficient increases with an increase in aqueous phase pH or CTAB concentration before reaching a saturated state. The existence of anions in the system can affect the extraction in several ways. For example, SO$_{4}^{2-}$ and PO$_{4}^{3-}$ ions can form hydrogen bonds with hyperoside and thereby improve the extraction significantly.CONCLUSION: This study proved that extraction using CTAB reverse micelles is a very effective method to separate hyperoside from Hypericum perforatum L. Under optimal conditions the extraction efficiency can reach 68.3% and the single‐strip‐extraction efficiency can reach 85.9%. The proportion of hyperoside among the flavonoids left in the strip‐extract can reach 93.3 wt%. This finding is very significant, because it demonstrates that reverse micellar extraction is a promising way to separate and purify materials with small molecules. Copyright © 2008 Society of Chemical Industry

Formation of monetite nanoparticles and nanofibers in reverse micelles

Reverse micelles solution of water and cyclohexane containing either cetyltrimethylammonium bromide (CTAB) or polyoxyethylene-8-dodecyl ether (C12E8) surfactants and n-pentanol as co-surfactant have been used as organized reaction microenvironments for monetite (dicalcium phosphate anhydrous, DCPA) precipitation. Well-crystallized monetite nanoparticles with various morphologies such as spheres, nanofibers and bundles of nanowires were obtained in CTAB reverse micelles solution. The molar ratio of water and surfactant (Wo) and the molar ratio of co-surfactant and surfactant (Po) have great influence on the structure and morphology of the final products. A generalized mechanism for the growth of monetite in reverse micelles is proposed, in which the interaction between the surfactant molecules and PO43− ions leads to the formation of a surfactant/CaHPO4 complex. It is because of this central complex that the further fusion with reactant ions containing reverse micelles will occur only in one direction. Changing the content of water and co-surfactant has great influence on the morphology of reverse micelles and on the interaction between the surfactant/CaHPO4 complex leading to a fine tuning of the morphology of products. By contrast, lacking of this interaction in the C12E8 system only tablet amorphous calcium phosphate can be formed.

Ultrafast dynamics of water in cationic micelles

The effect of confinement on the dynamical properties of liquid water is investigated for water enclosed in cationic reverse micelles. The authors performed mid-infrared ultrafast pump-probe spectroscopy on the OH-stretch vibration of isotopically diluted HDO in D(2)O in cetyltrimethylammonium bromide (CTAB) reverse micelles of various sizes. The authors observe that the surfactant counterions are inhomogeneously distributed throughout the reverse micelle, and that regions of extreme salinity occur near the interfacial Stern layer. The authors find that the water molecules in the core of the micelles show similar orientational dynamics as bulk water, and that water molecules in the counterion-rich interfacial region are much less mobile. An explicit comparison is made with the dynamics of water confined in anionic sodium bis(2-ethythexyl) sulfosuccinate (AOT) reverse micelles. The authors find that interfacial water in cationic CTAB reverse micelles has a higher orientational mobility than water in anionic AOT reverse micelles.

Enzymatic hydrolysis of microcrystalline cellulose in reverse micelles

The activities of cellulases from Trichoderma reesei entrapped in three types of reverse micelles have been investigated using microcrystalline cellulose as the substrate. The reverse micellar systems are formed by nonionic surfactant Triton X-100, anionic surfactant Aerosol OT (AOT), and cationic surfactant cetyltrimethyl ammonium bromide (CTAB) in organic solvent mediae, respectively. The influences of the molar ratio of water to surfactant ω 0, one of characteristic parameters of reverse micelles, and other environmental conditions including pH and temperature, on the enzymatic activity have been studied in these reverse micellar systems. The results obtained indicate that these three reverse micelles are more effective than aqueous systems for microcrystalline cellulose hydrolysis, and cellulases show “superactivity” in these reverse micelles compared with that in aqueous systems under the same pH and temperature conditions. The enzymatic activity decreases with the increase of ω 0 in both AOT and Triton X-100 reverse micellar systems, but reaches a maximum at ω 0 of 16.7 for CTAB reverse micelles. Temperature and pH also influence the cellulose hydrolysis process. The structural changes of cellulases in AOT reverse micelles have been measured by intrinsic fluorescence method and a possible explanation for the activity changes of cellulases has been proposed.

Effects of Nonpolar Solvents on the Solubilization of Pepsin into Bis(2-Ethylhexyl) Sodium Sulfosuccinate and Cetyltrimethylammonium Bromide Reverse Micelles

Solubilization of pepsin by bis(2-ethylhexyl) sodium sulfosuccinate (AOT) and cetyltrimethylammonium bromide (CTAB) reverse micelles has been studied at 20∘C. Isooctane, cyclohexane and hexane were used as solvents, and n-butanol, amyl alcohol and hexanol were used as cosurfactants for CTAB. AOT concentrations were varied from 50 to 500 mM and pepsin concentrations were varied from 2 to 10 mg-mL−1. At 250 mM, AOT can solubilize more than 85% of the Pepsin in each solvent. The effect of aqueous-phase pH on the solubilization of Pepsin has been studied from pH 1 to 8. The maximum solubilization of pepsin was observed below the isoelectric point (pI = 1.5) of the protein at pH 1.0 with 300 mM of AOT. The CTAB solutions were prepared by dissolving CTAB in isooctane with varying concentrations (0–100% v/v) of n-butanol, amyl alcohol or hexanol cosurfactants. It was found that 5% cosurfactant with 100 mM of CTAB was sufficient to solubilize more than 90% of the total pepsin. Pepsin solubilization by AOT reverse micelles increases with increasing polarizability and molar volume of the solvents.

Forward and backward extraction of BSA using mixed reverse micellar system of CTAB and alkyl halides

The forward and backward extraction of bovine serum albumin (BSA) has been studied using mixed reverse micellar systems of cetyl trimethyl ammonium bromide (CTAB) and alkyl halides (including 1-chlorobutane (R 4Cl), 1-bromooctane (R 8Br) and 1-iodobutane (R 4I)). The addition of alkyl halide R 8Br to 50 mmol/l CTAB/20% (v/v) hexanol/petroleum ether reverse micellar system could improve the BSA transfer from the aqueous phase to reverse micellar phase, while the addition of R 4Cl or R 4I had almost no effect on the BSA transfer. The mixed reverse micelles formed with CTAB and alkyl halides exhibit excellent backward extraction behavior for BSA. The mixed reverse micelles formed with CTAB and R 4I can realize the recovery of BSA effectively in a wide range of pH up to or higher than the isoelectric point of BSA. The mixed reverse micelles formed with CTAB and R 4Cl, R 8Br or R 4I can obviously enhance the BSA backward transfer at low ionic strength with addition of KBr or KCl as electrolyte. The mixed reverse micellar system indicates that it requires less time to reach the mass transfer equilibrium in comparison with the reverse micellar system with CTAB only. The mechanism of backward extraction proposed that with the addition of alkyl halides to CTAB reverse micelles, the hydrophobic interaction between the reverse micelles decreased.