Scorpion digestive lipase: kinetic study using monomolecular film technique.
Using the classical emulsified system and the monomolecular film technique, we compared the interfacial properties of the scorpion digestive lipase (SDL) with those of higher animals'. In the absence of bile slats, SDL does not hydrolyse efficiently pure tributyrin, as well as dicaprin films maintained at low surface pressure. The preincubation of bile salts with tributyrin seems to be a better substrate for SDL than the pure tributyrin. A kinetic study on the surface pressure dependency, stereospecificity and regioselectivity of SDL was performed using monomolecular films of either three dicaprin isomers or three pairs of didecanoyl-deoxyamino-O-methyl glycerol enantiomers (DDG) containing a single hydrolysable decanoyl ester bond. With all diacylglycerol isomers, SDL has a surface pressure threshold of about 15 m Nm(-1), below which enzymatic activity is undetectable. SDL seems to prefer vicinal ester groups of the diacylglycerol isomers, with preference for sn-1 position at both 15 and 23 m Nm(-1). Furthermore, the maximum SDL activity is measured with DDG having a primary ester bond (1,3DDG, SII). This shows that SDL has a preference for the sn-1 position of this diacylglycerol analogue. Moreover, this was in line with the fact that SDL is inactive on sn-2 position of both DDG isomers and a triacylglycerol. With diacylglycerol analogue isomers, SDL shows a preference for distal isomers contrary to what has been observed with diacylglycerol isomers. SDL interacts with egg-phosphatidyl choline (egg-PC) monomolecular films. The critical surface pressure value (13 m Nm(-1)) is comparable to those of pancreatic lipases.
Laboratoire de Biochimie et de Genie Enzymatique des Lipases, ENIS, BPW 3038 Sfax, Tunisia.
Nacim
N
Zouari
Adel
A
Sayari
Nabil
N
Miled
Robert
R
Verger
Youssef
Y
Gargouri
D000818:Animals; D001647:Bile Acids and Salts / Q000737:chemistry; D004075:Diglycerides / Q000737:chemistry; D004530:Egg Yolk / Q000737:chemistry; D005989:Glycerides / Q000737:chemistry; D043143:Hepatopancreas / Q000201:enzymology; D006868:Hydrolysis; D007700:Kinetics; D008049:Lipase / Q000737:chemistry*; D008567:Membranes, Artificial; D010713:Phosphatidylcholines / Q000737:chemistry; D013237:Stereoisomerism; D013379:Substrate Specificity; D013499:Surface Properties; D013997:Time Factors
D001647:Bile Acids and Salts; D004075:Diglycerides; D005989:Glycerides; D008567:Membranes, Artificial; D010713:Phosphatidylcholines; C512041:didecanoyl-deoxyamino-O-methyl glycerol; C044297:1,2-didecanoylglycerol; D008049:Lipase