Enzyme-Catalyzed Hydrolysis of Lipids in Immiscible Microdroplets Studied by Contained-Electrospray Ionization

Enzyme-catalyzed hydrolysis of lipids was monitored directly in immiscible microdroplet environments using contained-electrospray mass spectrometry. Aqueous solution of the lipase enzyme from Pseudomonas cepacia and the chloroform solution of the lipids were sprayed from separate capillaries, and the resultant droplets were merged within a reaction cavity that is included at the outlet of the contained-electrospray ionization source. By varying the length of the reaction cavity, the interaction time between the enzyme and its substrate was altered, enabling the quantification of reaction product as a function of time. Consequently, enhancement factors were estimated by comparing rate constants derived from the droplet experiment to rate constants calculated from solution-phase conditions. These experiments showed enhancement factors greater than 100 in favor of the droplet experiment. By using various lipid types, two possible mechanisms were identified to account for lipase reactivity in aerosols: in-droplet reactions for relatively highly soluble lipids and a droplet coalescence mechanism that allows interfacial reactions for the two immiscible systems.