Molecular mechanisms of fatty acids influence on bio-imprinted Rhizomucor miehei lipase towards 1,3-dioleoyl-2-palmitoyl glycerol synthesis

Four fatty acids (lauric acid (LA), myristic acid, palmitic acid, and stearic acid were employed as the template molecules to imprint Rhizomucor miehei lipase (RML) for the synthesis of 1,3-dioleoyl-2-palmitoylglycerol (OPO) by acidolysis reaction. Results indicated that the catalytic activity of bio-imprinted RML was higher than non-imprinted RML but decreased as the aliphatic chain length of fatty acids increased. The mechanism was studied through molecular dynamics simulation, and we found that fatty acids-imprinting affected the whole conformation, lid domain, and active site of RML, thus resulting in the diverse effects of fatty acids with different aliphatic chain lengths on the substrate accessibility, catalytic triad, and the stability of catalytic intermediate of bio-imprinted RML. Finally, LA-imprinted RML was utilized to catalyze OPO synthesis. Upon the optimal reaction conditions, C52 content and ( sn -2, P)/P value of OPO product reached 58% and 65%, respectively. This study would provide structure-function insights for designing efficient bio-catalysts and contribute to the development of bio-catalysts for food industry.