Crystallization and characterization of monoacylglycerol and diacylglycerol lipase from Penicillium camembertii.
A new lipase from Penicillium camembertii U-150, which is specific for monoacylglycerols and diacylglycerols, but not triacylglycerols, was purified as four active components using concanavalin-A-Sepharose column chromatography, crystallized in the form of needles, and its properties investigated. No significant difference was observed in substrate specificity, but molecular mass and other enzymatic properties, such as pH, heat stability and optimum pH and temperature, were clearly different between the unadsorbed and the three adsorbed components on concanavalin-A-Sepharose; the three adsorbed components were similar to each other and more stable than the unadsorbed component. On the other hand, after enzymatic removal of carbohydrates from the three adsorbed components, their enzymatic properties became similar to those of the unadsorbed component. The carbohydrates of this lipase contribute to the stability of the enzyme, but not to its enzyme activity. The amino acid compositions of the four components did not differ from each other, and tryptic mapping of the deglycosylated components and amino acid composition of the tryptic fragments were identical. The carbohydrate compositions of four intact components were, however, different from each other. All four components have the same polypeptide backbone and multiple forms of this lipase are due to the differences in composition of the carbohydrates bound in this lipase.
Isobe
K
K
Research and Development Division, Amano Pharmaceutical Co. Ltd., Nagoya, Japan.
Nokihara
K
K
Yamaguchi
S
S
Mase
T
T
Schmid
R D
RD
D000596:Amino Acids / Q000032:analysis; D050260:Carbohydrate Metabolism; D002851:Chromatography, High Pressure Liquid; D003460:Crystallization; D004952:Esters / Q000378:metabolism; D006031:Glycosylation; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007525:Isoelectric Focusing; D007526:Isoelectric Point; D008071:Lipoprotein Lipase / Q000302:isolation & purification / Q000378:metabolism*; D008970:Molecular Weight; D008994:Monoacylglycerol Lipases / Q000302:isolation & purification / Q000378:metabolism*; D010407:Penicillium / Q000201:enzymology*; D010449:Peptide Mapping; D013696:Temperature; D014357:Trypsin / Q000737:chemistry
D000596:Amino Acids; D004952:Esters; D008994:Monoacylglycerol Lipases; D008071:Lipoprotein Lipase; D014357:Trypsin