Partial purification of a lipolytic enzyme from Escherichia coli

• Published in: Canadian journal of biochemistry Vol. 56; no. 5; p. 319
• Main Authors: Proulx, P, Nantel, G, Baraff, G
• Format: Journal Article
• Language: English
• Published: Canada 01.05.1978
• Subjects: Escherichia coli - enzymology; Kinetics; Phospholipases - isolation & purification; Phospholipases - metabolism; Substrate Specificity
• ISSN: 0008-4018
• DOI: 10.1139/o78-049

An enzyme with phospholipase Al activity was purified some 500-fold from Escherichia coli cell homogenates. Lipase, phospholipase A2, and lysophospholipase copurified with phospholipase A1 and the four activities displayed similar susceptibility to heat treatment. The phospholipase A and lipase activities were recovered in a single band when partially purified preparations were subjected to SDS gel electrophoresis. Phospholipase, lysophospholipase, and lipase all required Ca2+ for activity. Phosphatidylcholine, phosphatidylethanolamine, and their lyso analogues were all hydrolysed at equivalent rates and these were substantially greater than the rate of methylpalmitate or tripalmitoylglycerol hydrolyses under similar incubation conditions. Evidence for a direct but slow hydrolysis of the ester at position 2 of phosphoglyceride was obtained; however, release of fatty acid from this position is mostly indirect involving acyl migration to position 1 and subsequent release of the translocated fatty acid. Escherichia coli, therefore, appears to possess a lipolytic enzyme of broad substrate specificity acting mainly at position 1 but also at position 2 of phosphoglycerides and on triacylglycerols and methyl fatty-acid esters.