Action of cutinase at the triolein-water interface. Characterisation of interfacial effects during lipid hydrolysis using the oil-drop tensiometer as a tool to study lipase kinetics.
Interfacial events during lipid hydrolysis by cutinase are described as measured with the oil-drop tensiometer. A linear relation between enzyme concentration and initial decrease of oil-water interface tension (gamma o/w) due to lipolytic activity was observed. The amount of hydrolysis products showed a non-linear relation with gamma o/w. Hydrolysis is linear with time, even when the area occupied by the fatty acid molecules exceeds the drop surface by a factor 7000. At pH 9.0, fatty acids were found to partition mainly in the oil phase. Formation of calcium soaps and ionization increase the impact of fatty acids on gamma o/w without affecting enzyme activity. The presence of fatty acids at the interface, added prior to cutinase, delayed hydrolysis effects on gamma o/w. Fatty acids in the water phase almost completely abolished adsorption effects on gamma o/w, when the concentration was over the critical micellar concentration (cmc).
Flipsen
J A
JA
Department of Enzymology and Protein Engineering, Utrecht University, The Netherlands.
van der Hijden
H T
HT
Egmond
M R
MR
Verheij
H M
HM
D055592:Biophysical Phenomena; D001703:Biophysics; D002265:Carboxylic Ester Hydrolases / Q000235:genetics / Q000378:metabolism*; D005227:Fatty Acids; D005670:Fusarium / Q000201:enzymology / Q000235:genetics; D006868:Hydrolysis; D007700:Kinetics; D008049:Lipase / Q000378:metabolism*; D008823:Micelles; D017354:Point Mutation; D013500:Surface Tension; D014304:Triolein; D014867:Water
D005227:Fatty Acids; D008823:Micelles; D014867:Water; D014304:Triolein; D002265:Carboxylic Ester Hydrolases; C019952:cutinase; D008049:Lipase