Improving an approach to lipase determination in enzyme preparations for the food industry based on a method of fatty acid production

The production of enzyme preparations remains to be a promising and permanently growing direction of modern biotechnology. The industrial production of the most widely used enzyme preparations contributes not only to improving the existing technologies, but also to developing fundamentally new processes. In this connection, lipases as part of the processes of partial or complete hydrolysis of oils and fats present significant research interest. Lipases find application in food processing and consumer goods manufacturing, as well as in agriculture, medicine, and analytical practice. In this study, we compare three methods based on the reaction of hydrolytic cleavage of ester bonds in a fat molecule followed by the release of free fatty acids. The optimum parameters providing for the highest production of fatty acids were determined. A substrate to obtain a more stable emulsion under a higher accuracy of measurements was experimentally selected. The established optimum conditions for a biocatalytic reaction of substrate hydrolysis (olive oil) using the conventional enzyme preparation “Lipase” included the temperature of 40 °С, the pH of 4.7, and the duration of 20 minutes. In order to confirm the validity of the data and parameters obtained, comparative studies were carried out to determine lipase activity in various objects of microbial origin. When varying the dosage of the enzyme preparation, the lipase activity was found to be within 5% of measurement accuracy. This error agrees well with the conventional methods of determining the activity of enzyme preparations for the food industry.

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