Application of reverse micelles for the extraction of proteins

Leser, Martin E.; Wei, G.; Luisi, Pier Luigi; Maestro, M.

doi:10.1016/0006-291x(86)90039-2

Cited by 103 publications

(71 citation statements)

References 8 publications

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“…2,5,[11][12][13][17][18][19]21 In this study, we also observed such a dependence for the extraction efficiency of Cyt c, as shown in Fig. 4.…”

Section: Dependence Of the Extraction Efficiency On The Electrolyte Csupporting

confidence: 78%

“…Since pioneering work by Luisi et al, [1][2][3] a great deal of interest has been focused on the solvent extraction of proteins or enzymes via reverse micelle formation. The reverse micelle extraction of proteins is useful for the separation and purification of proteins, 6 enzymatic reactions in organic solvents, 7,8 refolding of denatured proteins, 9,10 etc.…”

Section: Introductionmentioning

confidence: 99%

“…To understand the extraction mechanism, many efforts have been made to study the effects of various experimental conditions, which include the natures and concentrations of proteins, surfactants, and aqueous electrolytes and the pH of the aqueous phase. [2][3][4][5][6][11][12][13][14][15][16][17][18][19][20][21] It is generally recognized that the electrostatic interactions between a protein and surfactants play an important role in the formation of reverse micelles containing proteins. It has also been claimed that the electrostatic interactions of a protein with a bulk oil/water (O/W) interface play the dominant role in the inclusion of a protein in a reverse micelle at the O/W interface.…”

Section: Introductionmentioning

confidence: 99%

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Electrochemical Aspects of the Reverse Micelle Extraction of Proteins

Osakai

Shinohara

2008

ANAL. SCI.

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The mechanism of the solvent extraction of cytochrome c (Cyt c) via reverse micelle formation was studied from an electrochemical point of view. Potentiometric measurements showed that the Galvani potential difference of the oil/water (O/W) interface played a crucial role in the spontaneous extraction of Cyt c with bis(2-ethylhexyl)sulfosuccinate (AOT). However, the dependence of the extraction efficiency on the concentration of an aqueous electrolyte (KCl) could be explained not by the effect of the interfacial potential, but by the change in the interfacial tension (g). Electrocapillary measurements showed that the adsorption of AOT anions to the O/W interface resulted in a significant decrease of g in a higher potential range, where reverse micelles were formed. The bottom level of g in the higher potential range was increased with [KCl]. The lower extraction efficiency for higher [KCl]'s was elucidated by a "size exclusion effect". This was also supported by water-content measurements by the Karl Fisher method.

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“…2,5,[11][12][13][17][18][19]21 In this study, we also observed such a dependence for the extraction efficiency of Cyt c, as shown in Fig. 4.…”

Section: Dependence Of the Extraction Efficiency On The Electrolyte Csupporting

confidence: 78%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Electrochemical Aspects of the Reverse Micelle Extraction of Proteins

Osakai

Shinohara

2008

ANAL. SCI.

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“…(Microemulsions of smaller size are referred to as ''reverse micelles.'') For example, liquid-liquid extraction using w/o-MEs have been used to selectively isolate target proteins from aqueous solutions or solids (Leser and Luisi, 1990;Pires et al, 1996). In addition, w/o-MEs are ideally suited for hosting enzymatic reactions, especially those that involve difficultto-solubilize substrates, require low water content, or employ multiple biocatalysts (Luisi et al, 1988;Oldfield, 1994).…”

Section: Introductionmentioning

confidence: 99%

“…Luisi and co-workers determined that SPE was driven by either electrostatic interactions between biomolecule and surfactant head group or the affinity of the biomolecule for the w/o-ME's dispersed aqueous phase (Leser and Luisi, 1990). Whether electrostatic interactions or water affinity drove the extraction depended on the size of the w/o-MEs, or equivalently, the solution's water-AOT molar ratio, known as w o (Leser and Luisi, 1990).…”

Section: Introductionmentioning

confidence: 99%

Mechanism of protein extraction from the solid state by water-in-oil microemulsions

Hayes

1997

Biotechnol. Bioeng.

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The extraction of solid‐phase α‐chymotrypsin, bovine serum albumin (BSA), and lysozyme by water‐in‐oil microemulsion (w/o‐ME) solution containing Aerosol‐OT (AOT) was thoroughly examined as a means to maximize protein solubilization in organic solvent media. Protein extraction occurred simultaneously with the adsorption of water and AOT by the solid protein. Water and AOT were desorbed at nearly equal rates, suggesting that both materials were desorbed together as micreomulsions. The solubilization of protein increased linearly with the ratio of solid protein to extractant solution except at a high value of the ratio, where most protein‐containing microemulsions were desorbed. Based on our results, a mechanistic model was developed to describe the solid‐phase extraction procedure. First, microemulsions are desorbed from solution by the solid protein, resulting in the formation of a solid protein‐AOT‐water aggregate. Second, when a protein in the solid phase binds to a sufficient number of microemulsions, the resulting aggregate's increased hydrophobicity drives its solubilization into lipophilic solvent. Third, through the exchange of materials between the solubilized precipitate and the remaining microemulsions, protein‐containing w/o‐MEs are formed. The presence of adsorption is further indicated by an isotherm existing between the water, AOT, and protein content of the resulting solid phase for each protein. The driving force behind adsorption is either AOT‐protein interactions or the protein's affinity for microemulsion‐encapsulated water, depending on the properties of the protein and the size of the microemulsions, in agreement with the model of P. L. Luisi [Chimia, 44: 270–282 (1990)]. The second step of our model is mass transfer limited for the extraction of solid α‐chymotrypsin and BSA. The extraction of solid lysozyme was limited by the occurrence of an irreversible precipitation process. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 53: 583–593, 1997.

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Protein extraction using the sodium bis(2-ethylhexyl) phosphate (NaDEHP) reverse micellar system

Gülari

1996

Biotechnol. Bioeng.

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The reverse micellar system of sodium bis(2-ethylhexyl) phosphate (NaDEHP)/isooctane/brine was used for liquidliquid extraction of proteins. We investigated the solubilization of cytochrorne-c and 0-chymotrypsin into the NaDEHP reverse micellar phase by varying the pH and NaCl concentration in the aqueous phase. At neutral pH and relatively low ionic strength, the proteins are extracted into the micellar phase with high yield. By contacting the micellar phase with a divalent cation (e.g., Ca") aqueous solution, the reverse micelles are destabilized and release the protein molecules back into an aqueous solution for recovery. This method separates the proteins from the surfactant with very high overall efficiencies.

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Application of reverse micelles for the extraction of proteins

Cited by 103 publications

References 8 publications

Electrochemical Aspects of the Reverse Micelle Extraction of Proteins

Electrochemical Aspects of the Reverse Micelle Extraction of Proteins

Mechanism of protein extraction from the solid state by water-in-oil microemulsions

Protein extraction using the sodium bis(2-ethylhexyl) phosphate (NaDEHP) reverse micellar system

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