Monolayer techniques for studying lipase kinetics.

Abstract

Publisher Summary This chapter discusses the why lipid monolayers is used as lipase substrates for studying lipase kinetics. The monolayer is a suitable model system to study the enzymatic reactions carried out at the interfaces in a heterogeneous medium. There are at least five major reasons for using lipid monolayers as substrates for lipolytic enzymes. To use the monolayer technique for lipase kinetics, one necessary condition is that the substrate must be insoluble in the aqueous subphase and must form stable monomolecular films. The monolayer technique is ideally suited for studying the mode of action of lipolytic enzymes at interfaces using controlled mixtures of lipids. Two methods exist for forming mixed lipid monolayers at the air/water interface: either by spreading a mixture of water-insoluble lipids from a volatile organic solvent, or by injecting a micellar detergent solution into the aqueous subphase covered with preformed insoluble lipid monolayers.