Yarrowia Lipolytica LIP2 Lipase, a Potent Drug Candidate
for Enzyme Replacement Therapy in Pancreatic Exocrine Insufficiency
Frédéric Carrière, Ahmed Aloulou, Mathieu Schué, Delphine Puccinelli, Stéphane Milano, Chantal Delchambre, Yves Leblond, René Laugier, 
CNRS, Aix-Marseille Université, EIPL UMR 7282
Marseille, France

The LIP2 lipase from the yeast Yarrowia lipolytica (YLLIP2) was proposed as a good drug candidate for enzyme replacement therapy in patients with pancreatic exocrine insufficiency (PEI) [1,2], but its potency was never tested under physiological conditions. The specific activity on dietary triacylglycerides (TAG) and the stability of YLLIP2 were measured in vitro under conditions mimicking those prevailing in the gastrointestinal (GI) tract during the digestion of a meal. Various pH conditions were tested in the presence and absence of bile and pepsin in order to predict the possible action of YLLIP2 in vivo. Similar in vitro digestion experiments were carried out using recombinant gastric (rDGL) and pancreatic (rHPL) lipases, the digestive lipases normally present in the GI tract, in order to identify the lipases with the highest potency. YLLIP2 displayed the highest specific activity on test meal TAG in the 4 to 7 pH range where it was highly stable and poorly degraded by pepsin. At lower pH values (2-3), rDGL was the most active and stable lipase, and YLLIP2 was sensitive to pepsin hydrolysis after pH inactivation. The doses of YLLIP2 to be administered to PEI patients were deduced from YLLIP2 specific activity on meal TAG and compared to the doses of pancreatin currently given to these patients. These doses were then tested in pancreatic duct-ligated minipigs used as a model of PEI. 

[1]	Aloulou, A., Rodriguez, J.A., Puccinelli, D., Mouz, N., Leclaire, J., Leblond, Y., Carriere, F., Purification and biochemical characterization of the LIP2 lipase from Yarrowia lipolytica, Biochimica et Biophysica Acta. 2007, 1771, 228–237.
[2]	Aloulou, A., Puccinelli, D., De Caro, A., Leblond, Y., Carriere, F., A comparative study on two fungal lipases from Thermomyces lanuginosus and Yarrowia lipolytica shows the combined effects of detergents and pH on lipase adsorption and activity, Biochim Biophys Acta. 2007, 1771, 1446-1456.