Reverse Micellar Extraction and Precipitation of Lysozyme Using Sodium Di(2-ethylhexyl) Sulfosuccinate

Abstract

Sodium di(2-ethylhexyl) sulfosuccinate, referred to as Aerosol-OT or AOT, was used to remove lysozyme from an aqueous phase via reverse micellar extraction and precipitation method. For both methods, when the surfactant was in excess, a complete removal of lysozyme from the aqueous phase was obtained at the values of pH below the pI of lysozyme. However, for the reverse micellar method, a solubilization limit of lysozyme in the organic phase was observed, and a white precipitate was formed at the aqueous-organic interface. This observation suggested using AOT directly as a precipitating ligand. The lysozyme precipitated with AOT was fully recovered, with its original enzymatic activity, using acetone as a recovery solvent. A mechanism is suggested to explain the solubilization of lysozyme in an AOT reverse micellar system. It is shown that a direct precipitation method can be used with advantage instead of using the reverse micellar extraction method to recover lysozyme from an aqueous phase.

Protein separation by affinity extraction with reversed micelles of Span 85 modified with Cibacron Blue F3G-A

“…The results indicated that the complex was unstable in the micellar phase, or in other words, the CB-Span 85 conjugate solubility in the reversed micelles decreased upon the complex formation with the protein. Similar phenomenon was also reported in the extraction of lysozyme with AOT reversed micelles [32,33]. Once getting contacted with a high ionic strength stripper, the complex was Table 1 Total recovery yields (E t ) of lysozyme stripped from different volumes of the organic phase (V o ) with and without hexanol addition (3 vol dissociated gradually and a transparent two-phase system was recovered.…”

Ionic liquid-based aqueous two-phase extraction of selected proteins

“…On the other hand, these techniques are not easily scalable. Liquid-liquid extraction of proteins using reverse micellar phase was proposed as an alternative method [5]. Normally reverse micellar extraction involves the use of volatile organic solvents such as isooctane [6].…”

Reverse Micellar Extraction of Bromelain from Pineapple (Ananas comosusL. Merryl) Waste: Scale-up, Reverse Micelles Characterization and Mass Transfer Studies

“…Low interfacial tension, ease of scale-up, and potential for continuous operation are some of the advantages of reverse micellar extraction (RME) (2,3). Several reports on the application of RME for separation and purification of biomolecules are available for model systems (4)(5)(6)(7)(8). In recent years, a few studies on RME of biomolecules from natural=real systems have been reported (9)(10)(11)(12)(13)(14)(15).…”