Hydrolysis of Triglycerides in Cream Using Porcine Pancreatic Lipase

Abstract

The development of an enzyme-mediated reaction for the first-year general, organic, and biological chemistry (GOB) teaching laboratory is of significant importance so that the factors that affect enzyme-mediated reactions can be adequately demonstrated, reinforcing the lecture portion of the GOB course. Some of the factors that affect these types of reactions include, but are not limited to, enzyme concentration, substrate concentration, and temperature. The experiment reported herein utilizes porcine pancreatic lipase (PPL) to catalyze the hydrolysis of triglycerides within cream. The amount of time required to afford triglyceride hydrolysis was probed by altering the concentration of PPL and the temperature at which the reaction was performed. In addition, to provide the students with a hands-on demonstration of how bile salts aid in the digestion of triglycerides, various trials were performed to explore the effect of altering bile salt concentration. Finally, the data obtained from the enzyme concentration trials, the temperature trials, and the bile salt concentration trials were graphed to provide a visual representation of how triglyceride hydrolysis, mediated by PPL, is affected by these various factors.

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“…The increase in the temperature was found to accelerate the mobility of substrate and lipase, leading to the higher reaction rate of the hydrolysis process, as presented in Figure 1D (Chua et al 2012). The result from CRL and PPL was fairly consistent with previous reports, in which the temperature range for the hydrolysis process was from 35-40 °C (Freitas et al 2007;Robertson and Passaro 2020;Sharma 2013). Lipozyme TL 100L was observed to have the highest activity at 50 °C, which was ascribed to its inherent characteristic from Thermomyces lanuginosus.…”