Kinetic assay of human gastric lipase on short- and long-chain triacylglycerol emulsions.
Under optimal conditions, assay for pure human gastric lipase was carried out with short- and long-chain triacylglycerol emulsions. Maximal specific activities of 1160 and 620 U/mg were obtained with tributyrin and soybean emulsion, respectively. We observed that with a tributyrin substrate, bovine serum albumin or bile salts must be added before the addition of the enzyme in order to prevent its irreversible interfacial denaturation. With long-chain triacylglycerols as substrate, a decrease with time in the rate of hydrolysis was associated with release of protonated long-chain fatty acids. The inhibitory effect of protonated fatty acids was also observed using tributyrin at pH 3.0. These observations support the conclusion that human gastric lipase shows no intrinsic specificity for short-chain triacylglycerols and that its apparent specificity is modulated by pH and presence of amphiphile in the incubation medium. Our conclusions support the view that, in the human, gastric lipolysis may play an important role in long-chain fat digestion.
D001647:Bile Acids and Salts / Q000378:metabolism; D004655:Emulsions; D005217:Fat Emulsions, Intravenous / Q000378:metabolism; D005750:Gastric Juice / Q000201:enzymology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D008049:Lipase / Q000378:metabolism*; D008066:Lipolysis; D019301:Oleic Acid; D009829:Oleic Acids / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000378:metabolism; D014280:Triglycerides / Q000378:metabolism*
D001647:Bile Acids and Salts; D004655:Emulsions; D005217:Fat Emulsions, Intravenous; D009829:Oleic Acids; D014280:Triglycerides; D012710:Serum Albumin, Bovine; D019301:Oleic Acid; D008049:Lipase; C005830:tributyrin