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 (γO/W) due to lipolytic activity was observed. The amount of hydrolysis products showed a...

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Bibliographic Details
Published in:Chemistry and physics of lipids 1996-12, Vol.84 (2), p.105-115
Main Authors: Flipsen, J.A.C., van der Hijden, H.T.W.M., Egmond, M.R., Verheij, H.M.
Format: Article
Language:English
Subjects:
Biophysical Phenomena
Biophysics
Carboxylic Ester Hydrolases - genetics
Carboxylic Ester Hydrolases - metabolism
Cutinase
Fatty Acids
Fusarium - enzymology
Fusarium - genetics
Hydrolysis
Hydrolysis products
Kinetics
Lipase - metabolism
Micelles
Oil-drop
Oil-water interfacial tension
Oleate
Oleic acid
Point Mutation
Surface Tension
Triolein
Water
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Summary: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 (γO/W) due to lipolytic activity was observed. The amount of hydrolysis products showed a non-linear relation with γO/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 γO/W without affecting enzyme activity. The presence of fatty acids at the interface, added prior to cutinase, delayed hydrolysis effects on γO/W Fatty acids in the water phase almost completely abolished adsorption effects on γO/W when the concentration was over the critical micellar concentration (cmc).
ISSN:0009-3084
1873-2941
DOI:10.1016/S0009-3084(96)02627-8