All you need to know about extraction of enzymes using reverse micelles!

M.V.N. UMA MAHESH

Surfactants , Micelles and Reverse Micelles

Surface active agents , commonly designated as surfactants , are amphiphilic molecules containing nonpolar and polar parts capable of interacting with surfaces. An example is given by the anionic surfactant AOT{bis(2-ethylhexyl)sulfosuccinate .

The term micelle is used to describe the aggregates of amphiphilic molecules in aqueous solutions and refers to soluble aggregate, spontaneously and reversibly formed from amphiphiles or ions. The surfactant molecules' selfaggregation occurs only above the critical micelle concentration (CMC) which in practical sense means that the surfactant has to be added of amount exceeding its solubility

Reversed micelle solvents represent nanometer-sized aqueous droplets stabilized by surfactants inside the bulk organic solvents. The structure of a reverse micelle consists of a an aqueous microdomain facing the polar heads of the surfactant that surrounds this core interacting with the bulk organic solvent (non-polar) through hydrophobic chains

Understanding Reverse Micelles

Formation of reverse micelles. Reverse micelle shape and size. Reverse micelle interface. Organic solvent, co-solvent, co-surfactant.

A ternary system containing two immiscible liquids (ex. water and hydrocarbon)and an amphiphile form different types of micro emulsion systems out of which stable water-inoil micro emulsion represents the reversed miceller structure Its structure involves a nanometersized aqueous droplet embraced by the hydrophilic heads of the surfactant molecules and stabilized inside a bulk organic solvent through interactions with the surfactant chains.

At constant temperature, the aqueous droplet size is a function only of the parameter W0 (=mol[water] / mol[surfactant]), a measure of the hydration degree. The reversed micelle size is linearly proportional to the ratioW0 above the value needed for hydration of the surfactant polar headgroups. The radius of the aqueous core of the reversed micelles is given considering simple spherical geometry and assuming that all the surfactant is aggregated in a micelle form (Regalado et al., 1994):

Reverse Miceller Extraction of Enzymes

The efficient reversed micelle extraction cycle consists of two successive mass transport processes Forward Extraction: Solute solubilization into the reversed micelles characterized by high extent and selectivity . Back Extraction: Subsequent solute stripping of sufficient amount and quality.

Forward Extraction( Solubilization )

Factors affecting Forward Extraction

Surfactant and Protein effects -Surfactant concentration -Protein Size Electrostatic interactions -pH effect -Ionic Strength effect Hydrophobic Interactions

Back Extraction( Desolubilization )

Electrostatic interactions -pH effect -Ionic Strength effect Other effects -Alcohol -Temperature -Counter Ionic Detergent

A Model for Estimation