Journal of Lipid Research, Vol 20, 437-446, Copyright © 1979 by Lipid Research, Inc.
Enzyme-substrate interaction in lipid monolayers. I. Experimental conditions and fundamental kinetics
DG Dervichian and JP Barque
A systematic study has shown the importance of the different factors which
are concerned with the action of lipase on a substrate (1,3-
didecanoylglycerol). These consist of a) the process of adsorption of
lipase to the surface, b) the necessity of limited stirring to reach
equilibrium, and c) the persistence during the reaction process of the
enzyme molecules adsorbed on the monolayer. On the basis of this
preliminary investigation, a technique was established to analyze the
mechanism of lipase action with defined quantities of enzyme and lipid
segregated in the monolayer. Thus, the process of the reaction itself is
separated from the adsorption process, and it is demonstrated that the
quantity of substrate hydrolyzed per minute depends only on the quantity of
initially adsorbed lipase and not on the quantity of substrate or on the
surface concentration of the enzyme. An appropriate new definition of the
rate is consequently adopted.
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