A monolayer and bulk study on the kinetic behavior of Pseudomonas glumae lipase using synthetic pseudoglycerides
- 1 October 1991
- journal article
- Published by American Chemical Society (ACS) in Biochemistry
- Vol. 30 (41), 10034-10042
- https://doi.org/10.1021/bi00105a031
Abstract
A heat-stable lipase from Pseudomonas glumae was purified to homogeneity. Its positional and stereospecific properties were investigated and compared with those of the well-known porcine pancreatic lipase. The kinetic properties of both enzymes were determined by use of six isomeric synthetic pseudoglycerides all composed of a single hydrolyzable fatty acyl ester bond and two lipase-resistant groups: one acylamino and one ether function. Two enzyme assay techniques were applied: a detergent-free system, the monomolecular surface film technique, and the pH-stat technique using clear micellar solutions of substrate in the presence of Triton X-100. Regarding the cleavage of primary ester bonds, P. glumae lipase possesses no stereopreference. In contrast, a large stereopreference in favor of the R-isomer is found for the hydrolysis of secondary ester bonds. Secondary ester bonds are efficiently cleaved by the lipase, which makes it of potential interest for enzymatic synthetic purposes. For the hydrolysis of this R-isomer a correlation between the experimental catalytic turnover rate and the binding constant for micelles was observed. The kinetic data of P. glumae lipase have been analyzed in terms of the scooting and hopping models for the action of lipolytic enzymes [Upreti, G.C., & Jain, M.K. (1980) J. Membr. Biol. 55, 113-121]. The results presented in this study are best explained by assuming that glumae lipase leaves the interface after a limited number of catalytic cycles.Keywords
This publication has 26 references indexed in Scilit:
- Competitive inhibition of lipolytic enzymes. III. Some acylamino analogues of phospholipids are potent competitive inhibitors of porcine pancreatic phospholipase A2Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism, 1990
- Competitive inhibition of lipolytic enzymes. II. Preparation of ‘monoacylamino’ phospholipidsBiochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism, 1990
- Competitive inhibition of lipolytic enzymes. I. A kinetic model applicable to water-insoluble competitive inhibitorsBiochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism, 1990
- The kinetics of interfacial catalysis by phospholipase A2 and regulation of interfacial activation: hopping versus scootingBiochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism, 1989
- Kinetic characterization of Escherichia coli outer membrane phospholipase A using mixed detergent-lipid micellesBiochemistry, 1989
- Lipoprotein lipase moves rapidly between lipid dropletsFEBS Letters, 1983
- Release of arachidonate from diglyceride in human platelets requires the sequential action of a diglyceride lipase and a monoglyceride lipaseBiochemical and Biophysical Research Communications, 1981
- Action of phospholipase A2 on unmodified phosphatidylcholine bilayers: Organizational defects are preferred sites of actionThe Journal of Membrane Biology, 1980
- Lipolytic EnzymesPublished by American Chemical Society (ACS) ,1974
- l-GlycidolJournal of the American Chemical Society, 1942