Liquid-Liquid Extraction of Proteins with Reversed Micelles

Abstract

The recovery of proteins using reversed micelles is a liquid-liquid extraction process that has received increasing attention since proteins were shown to be solubilized in organic solvents with surfactants, maintaining their functional properties, and to be transferred between an aqueous solution and a reversed micellar organic phase. This article reviews the application of reversed micellar systems as a bioseparation technique for isolation and purification of proteins. The parameters that affect protein solubilization into the reversed micelles and the equilibrium and kinetics aspects that are involved in the extraction and back-extraction of proteins are discussed. Several examples are also described including the application of this technique for purification of recombinant proteins: cytochrome b 5 and a cutinase from Fusarium solani pisi.

Eco-friendly flow injection amperometric determination of tert -butylhydroquinone in edible oil samples using micelle-mediated reverse-phase dispersive liquid–liquid microextraction and an unmodified screen-printed carbon electrode

“…Therefore, the optimal surfactant concentration in biphasic extraction systems corresponds to the minimum level above the CMC required to achieve maximum transfer ), provides the highest extraction efficiency. 30,31 The effect of vortex time on extraction efficiency was evaluated over the range of 20 to 100 s (Fig. 3d).…”

Eco-friendly flow injection amperometric determination of tert -butylhydroquinone in edible oil samples using micelle-mediated reverse-phase dispersive liquid–liquid microextraction and an unmodified screen-printed carbon electrode

“…Therefore, the optimal surfactant concentration in biphasic extraction systems corresponds to the minimum level above the CMC required to achieve maximum transfer ), provides the highest extraction efficiency. 30,31 The effect of vortex time on extraction efficiency was evaluated over the range of 20 to 100 s (Fig. 3d).…”

Structural changes of a protein extract from apple with polyphenoloxidase activity obtained by cationic reversed micellar extraction induced by high-pressure carbon dioxide and thermosonication

“…At pH above its isoelectric point, PPO presents a net negative charge that favors electrostatic interactions with the cationic surfactant. In any case, it must be highlighted that not only electrostatic interactions play an important role on protein solubilization but also hydrophobic interactions 9 . Rocha et al .…”

“…Effect of ionic strength: It has been shown that the buffers employed in this work can supply enough of the electrolytes to achieve the minimum value for reversed micelles formation with no phase separation. In the literature 9 , it has been reported that high ionic strength values lead to small micelles formation and it could reduce the interactions between the hydrophilic biomolecules and the polar group of the surfactant molecule. Therefore, the effect of higher values of the ionic strength by adding increasing amounts of KCl to the extraction medium was studied to determine the maximum salt concentration that can be present in the extraction medium to avoid a decrease in the protein solubilization in the organic phase.…”

“…At this concentration, the protein solubilization in the organic phase decreased. At a certain value of the ionic strength in the aqueous phase, electrostatic interactions between protein and surfactant decreased due to the Debye screening effect caused by the presence of mobile charge carriers 9 . Certain ionic strength is needed to form reverse micelles and phase separation, in this work, when no KCl was added, a good performance was observed.…”

“…Certain ionic strength is needed to form reverse micelles and phase separation, in this work, when no KCl was added, a good performance was observed. This could be attributed to the use of a buffered aqueous solutions that can supply enough ionic strength 9 . Imm and Kim 7 obtained also the best results when no KCl was added to the extraction medium, with 43% of activity recovery and a value of 13 for the purification fold.…”

Eco-friendly flow injection amperometric determination of tert -butylhydroquinone in edible oil samples using micelle-mediated reverse-phase dispersive liquid–liquid microextraction and an unmodified screen-printed carbon electrode

“…Therefore, the optimal surfactant concentration in biphasic extraction systems corresponds to the minimum level above the CMC required to achieve maximum transfer ), provides the highest extraction efficiency. 30,31 The effect of vortex time on extraction efficiency was evaluated over the range of 20 to 100 s (Fig. 3d).…”

Structural changes of a protein extract from apple with polyphenoloxidase activity obtained by cationic reversed micellar extraction induced by high-pressure carbon dioxide and thermosonication

“…At pH above its isoelectric point, PPO presents a net negative charge that favors electrostatic interactions with the cationic surfactant. In any case, it must be highlighted that not only electrostatic interactions play an important role on protein solubilization but also hydrophobic interactions 9 . Rocha et al .…”

“…Effect of ionic strength: It has been shown that the buffers employed in this work can supply enough of the electrolytes to achieve the minimum value for reversed micelles formation with no phase separation. In the literature 9 , it has been reported that high ionic strength values lead to small micelles formation and it could reduce the interactions between the hydrophilic biomolecules and the polar group of the surfactant molecule. Therefore, the effect of higher values of the ionic strength by adding increasing amounts of KCl to the extraction medium was studied to determine the maximum salt concentration that can be present in the extraction medium to avoid a decrease in the protein solubilization in the organic phase.…”

“…At this concentration, the protein solubilization in the organic phase decreased. At a certain value of the ionic strength in the aqueous phase, electrostatic interactions between protein and surfactant decreased due to the Debye screening effect caused by the presence of mobile charge carriers 9 . Certain ionic strength is needed to form reverse micelles and phase separation, in this work, when no KCl was added, a good performance was observed.…”

“…Certain ionic strength is needed to form reverse micelles and phase separation, in this work, when no KCl was added, a good performance was observed. This could be attributed to the use of a buffered aqueous solutions that can supply enough ionic strength 9 . Imm and Kim 7 obtained also the best results when no KCl was added to the extraction medium, with 43% of activity recovery and a value of 13 for the purification fold.…”

Eco-friendly flow injection amperometric determination of tert -butylhydroquinone in edible oil samples using micelle-mediated reverse-phase dispersive liquid–liquid microextraction and an unmodified screen-printed carbon electrode

“…Therefore, the optimal surfactant concentration in biphasic extraction systems corresponds to the minimum level above the CMC required to achieve maximum transfer ), provides the highest extraction efficiency. 30,31 The effect of vortex time on extraction efficiency was evaluated over the range of 20 to 100 s (Fig. 3d).…”

Structural changes of a protein extract from apple with polyphenoloxidase activity obtained by cationic reversed micellar extraction induced by high-pressure carbon dioxide and thermosonication

“…At pH above its isoelectric point, PPO presents a net negative charge that favors electrostatic interactions with the cationic surfactant. In any case, it must be highlighted that not only electrostatic interactions play an important role on protein solubilization but also hydrophobic interactions 9 . Rocha et al .…”

“…Effect of ionic strength: It has been shown that the buffers employed in this work can supply enough of the electrolytes to achieve the minimum value for reversed micelles formation with no phase separation. In the literature 9 , it has been reported that high ionic strength values lead to small micelles formation and it could reduce the interactions between the hydrophilic biomolecules and the polar group of the surfactant molecule. Therefore, the effect of higher values of the ionic strength by adding increasing amounts of KCl to the extraction medium was studied to determine the maximum salt concentration that can be present in the extraction medium to avoid a decrease in the protein solubilization in the organic phase.…”

“…At this concentration, the protein solubilization in the organic phase decreased. At a certain value of the ionic strength in the aqueous phase, electrostatic interactions between protein and surfactant decreased due to the Debye screening effect caused by the presence of mobile charge carriers 9 . Certain ionic strength is needed to form reverse micelles and phase separation, in this work, when no KCl was added, a good performance was observed.…”

“…Certain ionic strength is needed to form reverse micelles and phase separation, in this work, when no KCl was added, a good performance was observed. This could be attributed to the use of a buffered aqueous solutions that can supply enough ionic strength 9 . Imm and Kim 7 obtained also the best results when no KCl was added to the extraction medium, with 43% of activity recovery and a value of 13 for the purification fold.…”