Action of Humicola lanuginosa lipase on long-chain lipid substrates: 1. Hydrolysis of monoolein monolayers

The hydrolysis of 1-monoolein (MO) monomolecular films by Humicola lanuginosa lipase (HLL) was followed by measuring the simultaneous decrease with time in the film area and the surface potential using a ‘zero order trough’ at constant surface pressure (Verger and de Haas, Chem. Phys. Lipids 10 (1973) 127). The decrease with time in the film area reflects both the reduction in the area per molecule as well as the solubilization of the substrate and the product molecules during the transformation of the substrate MO into product of oleic acid (OA). The surface potential changes were interpreted as the results of the accumulation at the interface of negatively charged OA. The surface concentration of OA transiently present at the interface was determined by the surface pressure and the surface potential measurements on the basis of a developed kinetic model. In the proposed model we have taken into account the product and substrate solubilization rates in the presence of β-cyclodextrin (β-CD) as well as the flux supplied progressively by the moving barrier from the reservoir to the reaction compartment in order to keep the constant surface pressure. Values of the global kinetic constant Q_m were obtained. The selective lipolytic product acceptor, β-CD, accelerated considerably the hydrolytic process.