Abstract
Pancreatic lipase related-protein 2 (PLRP2) exhibits remarkable galactolipase and phospholipase A1 activities, which depends greatly on the supramolecular organization of the substrates and the presence of surfactant molecules such as bile salts. It is responsible for the effective hydrolysis of the main lipids composing plant photosynthetic membranes, which represent an excellent source of omega-3 for human nutrition. The objective of the study was to understand the modulation of the adsorption mechanisms and enzymatic activity of Guinea pig PLRP2 (gPLRP2), by the physical environment of the enzyme and physical state of its substrate at the nanoscopic and microscopic scales. Langmuir monolayers were used to reproduce homogeneous and heterogeneous photosynthetic model membranes containing galactolipid, and/or phospholipid, and/or phytosterol, presenting uncharged or charged interfaces. The same lipid mixtures were also used to form micrometric liposomes, and their digestion kinetics by gPLRP2 were investigated in the presence or absence of bile salts (NaTDC) during static in vitro, so called “bulk”, digestion. The enzymatic activity of gPLRP2 onto mixed monolayer of galactolipids and/or phospholipids, and/or phytosterols was characterized with an optimum activity at 15 mN/m, in the absence of bile salts. Additionally, gPLRP2 showed enhanced adsorption onto biomimetic model monolayer containing negatively charged lipids, but the compositional complexity in these systems induced a lag phase before the initiation of lipolysis, and a slowdown in enzymatic activity. In bulk, no enzymatic activity could be demonstrated on GL-based liposomes in the absence of bile salts, probably due to the high lateral pressure of the lipid bilayers. In the presence of NaTDC (4 mM), however, gPLRP2 showed both high galactolipase and moderate phospholipase A1 activities on liposomes, probably due to a decrease in packing and lateral pressure upon NaTDC adsorption, and subsequent disruption of liposomes. The understanding of PLRP2 enzymatic activity at the level of natural plant-based assemblies open the way to new strategies to vectorize bioaccessible omega-3 and rebalance human and animal diets.