title: Purification and substrate specificity of Staphylococcus hyicus lipase.
abstract: The Staphylococcus hyicus lipase gene has been cloned and expressed in Staphylococcus carnosus. From the latter organism the enzyme was secreted into the medium as a protein with an apparent molecular mass of 86 kDa. This protein was purified, and the amino-terminal sequence showed that the primary gene product was indeed cleaved at the proposed signal peptide cleavage site. The protein was purified from large-scale preparations after tryptic digestion. This limited proteolysis reduced the molecular mass to 46 kDa and increased the specific activity about 3-fold. Although the enzyme had a low specific activity in the absence of divalent cations, the activity increased about 40-fold in the presence of Sr2+ or Ca2+ ions. The purified lipase has a broad substrate specificity. The acyl chains were removed from the primary and secondary positions of natural neutral glycerides and from a variety of synthetic glyceride analogues. Thus triglycerides were fully hydrolyzed to free fatty acid and glycerol. The enzyme hydrolyzed naturally occurring phosphatidylcholines, their synthetic short-chain analogues, and lysophospholipids to free fatty acids and water-soluble products. The enzyme had a 2-fold higher activity on micelles of short-chain D-lecithins than on micelles composed of the L-isomers. Thus the enzyme from S. hyicus has lipase activity and also high phospholipase A and lysophospholipase activity.
authors.affiliation: Department of Biochemistry, State University of Utrecht, The Netherlands.
authors.forename: M G
authors.initials: MG
authors.lastname: van Oort
authors.forename: A M
authors.initials: AM
authors.lastname: Deveer
authors.forename: R
authors.initials: R
authors.lastname: Dijkman
authors.forename: M L
authors.initials: ML
authors.lastname: Tjeenk
authors.forename: H M
authors.initials: HM
authors.lastname: Verheij
authors.forename: G H
authors.initials: GH
authors.lastname: de Haas
authors.forename: E
authors.initials: E
authors.lastname: Wenzig
authors.forename: F
authors.initials: F
authors.lastname: Götz
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D003001:Cloning, Molecular; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008049:Lipase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D050356:Lipid Metabolism; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010194:Pancreatin / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D013210:Staphylococcus / Q000201:enzymology* / Q000235:genetics; D013324:Strontium / Q000494:pharmacology; D013379:Substrate Specificity; D013552:Swine; D014280:Triglycerides / Q000494:pharmacology
chemical_list: D010743:Phospholipids; D014280:Triglycerides; D010194:Pancreatin; D008049:Lipase; C005830:tributyrin; D002118:Calcium; D013324:Strontium