Manipulating The Environment Of a Single Lipase Molecule

Lipase are interfacial enzymes with attractive applications. Their activity is greatly enhanced in the presence of a hydrophobic surface, a process called interfacial activation. However, the kinetics of this behavior is not yet fully understood. We measured this kinetics of a lipase from Thermomyces Lanuginose at single enzyme level. We utilized single vesicle arrays as a novel biocompatible scaffold to immobilize enzymes and as an interface to study the effect of the enzyme's binding to the membrane on the observed activity. We used organic polymers to vary the accessibility of the enzyme molecule to the bilayer. By this kind of direct control of the microenviroment of the enzyme, we quantified the activity of enzyme at different degrees of accessibility to the bilayer. Therefore, we gained a clear insight of interfacial activation of lipase and we are currently using this platform to quantify the influence of additional parameters, such as size of vesicles and lipid composition, on enzyme's catalytic behavior.