Purification and biochemical characterization of Yarrowia lipolytica LIP2, a lipase of medical interest for the treatment of pancreatic exocrine insufficiency

Abstract

The LIP2 lipase from the yeast Y. lipolytica (YLLIP2) was obtained from two genetically modified strains with multi‐copies of the lip2 gene and further purified using gel filtration and cation exchange chromatography. Four YLLIP2 isoforms were identified and subjected to N‐terminal amino‐acid sequencing and mass spectrometry analysis. These isoforms differed in their glycosylation patterns and their molecular masses ranged from 36,874 to 38,481 Da, whereas the polypeptide mass was 33,385 Da. YLLIP2 substrate specificity was investigated using short, medium and long chain triglyceride (TG) at various pH and bile salt (BS) concentrations, and compared with those of human digestive lipases. YLLIP2 was not inhibited by bile salts with any of the substrate tested, and maximum specific activities were found to be 10,760 IU/mg on tributyrin, 16,920 IU/mg on trioctanoin and 12,260 IU/mg on olive oil at pH 6.0. YLLIP2 possesses the same pattern of regioselectivity as human pancreatic lipase, generating 2‐MG and free fatty acids, the lipolysis products absorbed at the intestinal level. YLLIP2 was found to be fairly stable and still active on long chain TG (1590 IU/mg) at pH 4.0, in the presence of BS. YLLIP2 is therefore tailored to act optimally under the physiological conditions pertaining in the gastro‐intestinal tract, and might be an ideal candidate for enzyme replacement therapy for pancreatic exocrine insufficiency.

Characterization of all the lipolytic activities in pancreatin and comparison with porcine and human pancreatic juices

“…Recombinant human pancreatic lipase-related protein 2 (rHPLRP2), guinea pig pancreatic lipase-related protein 2 (rGPLRP2) and Yarrowia lipolytica LIP2 lipase (YLLIP2) were produced in the yeast Pichia pastoris and purified as described previously in [23,24], [25] and [26], respectively. Native porcine pancreatic lipase (PPL) and colipase were purified to homogeneity from porcine pancreas according to methods previously described [27,28].…”

Pancreatic Enzyme Replacement Therapy in hyperglycemic Male Rats

“…Aloulou et al, (14) reported that replacing all functions of these lipases with a single enzyme may be best choice in diabetic rats that suffering from malabsorption of fat.…”

“…because the drug that were given play us replacement therapy for exocrine pancreatic enzyme and not treated the destruction and necrosis of beta cells that mean remaining the lack of insulin production and remain diseased of diabetes mellitus. (14).…”

Supplementation of Bile Acids and Lipase in Broiler Diets for Better Nutrient Utilization and Performance: Potential Effects and Future Implications – A Review

“…The lipase from YL is an ideal candidate for enzyme replacement therapy due to its unique biochemical properties: It shows the highest activity at low pH values and is not repressed by bile salts. The YL belongs to the same gene family as Thermomyces lanuginosus lipase, a well-known lipase with many applications in the field of detergents and biotechnological processes (Aloulou et al, 2007). Although these two lipases show a high sequence identity of 30.3%, they have relatively different biochemical properties.…”