title: Enzyme activity and structural dynamics linked to micelle formation: a fluorescence anisotropy and ESR study.
abstract: Activities of the enzymes, protease subtilisin and horse radish peroxidase (HRP) have been increased 50 and 40%, respectively, in the presence of the nonionic surfactant, alkyl polyglucoside, compared with the activities in buffer alone. This enzyme hyperactivity reaches a peak at 3.0 mm of surfactant. Investigation into the structure of surfactant aggregates indicates "giant" micelle superstructures at this range of surfactant concentration of 1.7 μm in diameter--dramatically decreasing to 60 and 70 nm at higher surfactant concentrations, while surface tension measurements indicate two critical micelle concentration inflection points at 0.2 and 5.0 mm, which suggests transitions in micelle structure with respect to concentration. Furthermore, electron spin resonance (ESR) indicates that the micelles in first critical micelle concentration regime are loosely packed relative to the second aggregate phase. We hypothesize that this loose packing results in diminished hydration shell repulsion between the micelles, leading to the large, micrometer-sized aggregates. We further hypothesize that it is the interaction with these loosely packed micelles that affects the flexibility of the HRP and protease enzyme structure. Time-resolved fluorescence anisotropy of subtilisin in Brij-30 indicates increasing flexibility of catalytic active site with surfactant concentration. This is correlated with an increase in enzymatic activity.
authors.affiliation: Earth and Environmental Engineering, Columbia University, New York, NY.
authors.forename: Michael
authors.initials: M
authors.lastname: Chin
authors.forename: Ponisseril
authors.initials: P
authors.lastname: Somasundaran
mesh_terms: D004578:Electron Spin Resonance Spectroscopy / Q000379:methods*; D005454:Fluorescence Polarization / Q000379:methods*; D006735:Horseradish Peroxidase / Q000737:chemistry / Q000378:metabolism*; D008823:Micelles; D011487:Protein Conformation; D020860:Subtilisin / Q000737:chemistry / Q000378:metabolism*
chemical_list: D008823:Micelles; D006735:Horseradish Peroxidase; D020860:Subtilisin