The effects of pH and ionic strength on the partitioning of four proteins in reverse micelle systems

Four proteins with different physicochemical properties have been partitioned in reversed micelle systems: thaumatin, ribonuclease A, soybean trypsin inhibitor, and α‐lactalbumin. The organic phase was formed by sodium salt (AOT) in isooctane, and the aqueous phase contained KCl, KBr, MgCl2, or NaCl. Aqueous phase pH was varied between 2 and 13 and ionic strength from 0.1 to 1.0 M. Small changes in pH [around the isoelecric point (pl)] were found to influence the solubilization of ribonuclease A and trypsin inhibitor, but for thaumatin the pH change necessary to affect partition was much greater as a consequence of the difference in net charge (titration curves) of these protein molecules as pH changes. The type of ions present in the system was also a determining factor for partition; the larger ions (K+) produced more electrostatic screening and hence less protein solubilization than the smaller ions (Na+). With changes in ionic strength surface hydrophobicity was a dominant factor affecting solubilization of thaumatin in NaCl‐containing systems at high pH. Charge distribution and hydrophobicity are thought to be important parameters when partitioning the protein α‐lactalbumin. © 1994 John Wiley & Sons, Inc.