Lysophospholipase of Escherichia coli.
A lysophospholipase from Escherichia coli cells was purified about 1,500-fold to near homogeneity by extraction with Tris-HCl buffer, streptomycin treatment, (NH4)2SO4 fractionation, column chromatographies on Sephadex G-200, DEAE-cellulose and hydroxylapatite-cellulose, and polyacrylamide gel electrophoresis. The final preparation had a molecular weight of 39,500 plus or minus 500. The enzyme hydrolyzes 1-acylglycerylphosphorylethanolamine, 2-acylglycerylphosphorylethanoiamine, and 1-acylglycerylphosphorylglycerol, but does not attack diacylphospholipids with long chain fatty acids, such as phosphatidylethanolamine and phosphatidylglycerol. The enzyme does not show any esterase activity against p-nitrophenyl acetate or palmitate. Although it does not hydrolyze triacylglycerol or diacylglycerol, it hydrolyzes 1-acylglycerol at almost the same rate as 1-acyl-sn-glycerol-3-phosphorylethanolamine. Results indicated that the acyl-hydrolyzing activities toward monoacyl-glycerylphosphorylethanolamine and monoacylglycerol belong to the same enzyme. In general, acidic and nonionic detergents inhibited the reaction. This lysophospholipase preparation hydrolyzes the monomolecular and micellar forms of lysophospholipids as well as of monoacylglycerol. The monomolecular and micellar forms of Triton X-100 both inhibited the hydrolyses of lysophospholipids and monoacylglycerol.
D002250:Carbon Radioisotopes; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology; D004983:Ethanolamines / Q000378:metabolism; D005994:Glycerophosphates / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008244:Lysophosphatidylcholines / Q000378:metabolism*; D008970:Molecular Weight; D010714:Phosphatidylethanolamines / Q000378:metabolism; D010715:Phosphatidylglycerols / Q000378:metabolism; D010740:Phospholipases / Q000302:isolation & purification* / Q000378:metabolism; D011092:Polyethylene Glycols / Q000494:pharmacology; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D013307:Streptomycin / Q000494:pharmacology; D013696:Temperature; D014357:Trypsin / Q000494:pharmacology
D002250:Carbon Radioisotopes; D004983:Ethanolamines; D005994:Glycerophosphates; D008244:Lysophosphatidylcholines; D010714:Phosphatidylethanolamines; D010715:Phosphatidylglycerols; D012967:Sodium Dodecyl Sulfate; D011092:Polyethylene Glycols; D010740:Phospholipases; D014357:Trypsin; D013307:Streptomycin