Relation between micellar structure of model bile and activity of esterase.
In a model bile solution composed of lecithin (L)-bile salt (B), the solubilization of lipid and the accessibility of enzyme to the lipid were examined by observation of EPR spectra and measurement of enzyme activity. The lifetime of the spin probe in the micellar phase was estimated to be approx. 1 microsecond by means of line shape analysis. Both population and lifetime increased with temperature and the molar ratio of lecithin to bile salt (L/B). The EPR data indicated that simple micelle of bile salt, mixed disk micelle of bile salt-lecithin, and multi-lamellar mixed disk micelle can exist in a model bile solution, depending on the L/B molar ratio across a range from 0 to 1.5. The maximal power of the mixed disk micelle to solubilize cholesteryl ester in the model bile at a L/B molar ratio of 1:1 was confirmed by EPR measurement of cholesteryl 12-DOXYL-stearate. Observation of the enzyme activity on a mixture of model bile and substrate at 37 degrees C revealed selective accessibility of cholesterol esterase (bovine pancreas) to mixed disk micelle, of cholesterol oxidase (Streptomyces cinnamomeus) to both simple and mixed disk micelle, and of pancreatic lipase (porcine pancreas) to both simple micelle and an oil droplet of substrate. The temperature-dependent activity of cholesterol oxidase to cholesterol in mixed disk micelle can be explained in terms of mesomorphic phase transition of lecithin side chains followed with fluidity of liquid crystal phase. Regarding phospholipase C from Bacillus cereus, though the selective accessibility to the micelles was not observed at 37 degrees C, a decrease in activity for mixed disk micelle could be found at lower temperatures.
Uematsu
S
S
School of Pharmaceutical Science, University of Shizuoka, Japan.
Uchida
T
T
Kinoshita
A
A
Kimura
F
F
Akahori
Y
Y
D001646:Bile / Q000737:chemistry*; D002788:Cholesterol Esters / Q000737:chemistry* / Q000378:metabolism; D002793:Cholic Acids / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D008823:Micelles; D008956:Models, Chemical; D002787:Sterol Esterase / Q000378:metabolism*; D013696:Temperature; D013816:Thermodynamics; D010738:Type C Phospholipases / Q000378:metabolism
D002788:Cholesterol Esters; D002793:Cholic Acids; D008823:Micelles; D002787:Sterol Esterase; D010738:Type C Phospholipases