MECHANISMS OF PROTEIN SOLUBILIZATION IN REVERSE MICELLES

Solubilization properties of alpha-chymotrypsin and alcohol dehydrogenase (LADH) in reverse micelles are reported for three different solubilization techniques. The solubilization properties for these two proteins depend on the method used for protein addition. The addition of a dry protein powder to a reverse-micelle-containing organic phase does not appreciably solubilize the protein until the diameter of the reverse micelle is similar to that of the protein. However, when an aqueous protein solution is injected into an organic phase, protein solubilization is not strongly dependent on micelle size. For chymotrypsin, multiple protein occupancy occurs at large micelle sizes, with as many as 11 chymotrypsin molecules solubilized in one reverse micelle. The solubilization of chymotrypsin using a phase-transfer technique with a positively charged surfactant follows the expected trend based on protein-surfactant electrostatic interactions. When a negatively charged surfactant is used for phase transfer, at low pH the solubilization data do not fit this electrostatic interaction mechanism. In this case, protein-surfactant aggregation may be occurring at the aqueous-organic interface.