Lipolysis of natural long chain and synthetic medium chain galactolipids by pancreatic lipase-related protein 2.
Monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG) are the most abundant lipids in nature, mainly as important components of plant leaves and chloroplast membranes. Pancreatic lipase-related protein 2 (PLRP2) was previously found to express galactolipase activity, and it is assumed to be the main enzyme involved in the digestion of these common vegetable lipids in the gastrointestinal tract. Most of the previous in vitro studies were however performed with medium chain synthetic galactolipids as substrates. It was shown here that recombinant guinea pig (Cavia porcellus) as well as human PLRP2 hydrolyzed at high rates natural DGDG and MGDG extracted from spinach leaves. Their specific activities were estimated by combining the pH-stat technique, thin layer chromatography coupled to scanning densitometry and gas chromatography. The optimum assay conditions for hydrolysis of these natural long chain galactolipids were investigated and the optimum bile salt to substrate ratio was found to be different from that established with synthetic medium chains MGDG and DGDG. Nevertheless the length of acyl chains and the nature of the galactosyl polar head of the galactolipid did not have major effects on the specific activities of PLRP2, which were found to be very high on both medium chain [1786+/-100 to 5420+/-85U/mg] and long chain [1756+/-208 to 4167+/-167U/mg] galactolipids. Fatty acid composition analysis of natural MGDG, DGDG and their lipolysis products revealed that PLRP2 only hydrolyzed one ester bond at the sn-1 position of galactolipids. PLRP2 might be used to produce lipid and free fatty acid fractions enriched in either 16:3 n-3 or 18:3 n-3 fatty acids, both found at high levels in galactolipids.
Amara
Sawsan
S
CNRS-Aix-Marseille Université-Enzymologie Interfaciale et Physiologie de la Lipolyse-UPR 9025, 31 Chemin Joseph Aiguier, 13402 Marseille Cedex 20, France.
Barouh
Nathalie
N
Lecomte
Jérôme
J
Lafont
Dominique
D
Robert
Sylvie
S
Villeneuve
Pierre
P
De Caro
Alain
A
Carrière
Frédéric
F
D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism; D002849:Chromatography, Gas; D002855:Chromatography, Thin Layer; D057075:Enzyme Assays; D005227:Fatty Acids / Q000378:metabolism; D038983:Galactolipids / Q000302:isolation & purification / Q000378:metabolism*; D006168:Guinea Pigs; D006801:Humans; D006868:Hydrolysis; D008049:Lipase / Q000378:metabolism*; D008066:Lipolysis; D018724:Spinacia oleracea / Q000737:chemistry / Q000378:metabolism
D005227:Fatty Acids; D038983:Galactolipids; C007388:digalactosyldiacylglycerol; C009909:monogalactosyldiacylglycerol; D002265:Carboxylic Ester Hydrolases; C019262:galactolipase; D008049:Lipase; C075988:pancreatic lipase related protein 2