Reverse Micelle Systems Composed of Water, Triton X-100, and Phospholipids in Organic Solvents: 1. Phase Boundary Titrations and Dynamic Light Scattering Analysis

Abstract

Enzymes entrapped in systems formed with water, phospholipids, toluene, and Triton X-100 show a catalytic activity that is much lower and a thermostability that is much higher than that observed in totally aqueous systems or in other types of reverse micelles. By phase boundary titrations and dynamic light scattering, this work characterizes reverse micelle systems formed in either toluene or propylbenzene with Triton X-100 and water. Four regions with distinct structural features were encountered. Up to one molecule of water per one Triton X-100 molecule, the system was transparent; light scattering measurements of this region indicated that water hydrated Triton X-100 monomers. A turbid region was formed as water content was increased to water:Triton X-100 ratios of 7.6 in toluene and 4.2 in propylbenzene. This thermodynamically unstable region was formed by large polydisperse structures. Transparent systems containing small size (27–150 Å) thermodynamically stable reverse micelles were formed when the ratio of water to Triton X-100 molecules in the reverse micelle was in the range of 7.6 to 26.8 in toluene and 4.2 to 15.1 in propylbenzene. In this region, micellar size increased with water content. Water concentrations higher than the latter values resulted in phase separation. A similar titration of the aforementioned systems in the presence of phospholipids revealed that in the first region of transparency up to 10 molecules of water hydrated a phospholipid molecule. The inclusion of phospholipids to the Triton X-100 systems caused a displacement of the boundaries of the second region of transparency toward higher water contents.