Enzymatic lipid removal from surfaces—lipid desorption by a pH-induced “electrostatic explosion”

Abstract

Removal of lipidic molecules from surfaces can be accomplished using detergents containing lipases. Surface cleaning is usually performed under alkaline conditions due to increased solubility of the hydrolysis products, especially free fatty acids. This paper shows that removal of a triacylglycerol film from a surface can be dramatically enhanced in a sequential system where pH is shifted to alkaline conditions after an initial lipolytic reaction period at or below neutral pH. Data from three different biophysical techniques, attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR), quartz crystal microbalance with dissipation monitoring (QCM-D), and total internal reflection fluorescence spectroscopy (TIRF) clearly show the effects of such cleaning procedure. Initially the reaction is carried out at pH below the p K a value of the fatty acids formed upon triacylglycerol hydrolysis, and the protonated fatty acids accumulate in the film. The mechanism of lipid removal, induced by increasing pH to a value above the fatty acid p K a, is explained by a burst caused by electrostatic repulsion between rapidly ionised fatty acids, i.e. by an “electrostatic explosion”. Performing the initial hydrolysis at pH 6 and the subsequent rinse at pH 10, using triolein as model substrate, lipid removal from surfaces by both commercial detergent lipases and non-commercial lipases was significantly improved compared to a reaction at constant pH 10.