title: Cloning and characterization of Photobacterium damselae ssp. piscicida phospholipase: an enzyme that shows haemolytic activity.
abstract: A phospholipase gene of Photobacterium damselae ssp. piscicida (ppp) was cloned from a genomic library and its nucleotide sequence was determined. The open reading frame consisted of 1218 bp encoding a protein of 405 amino acids with a predicted molecular mass of 46 kDa. The PPP had identities (53-55%) with phospholipase and haemolysin of Vibrio spp., while it showed low identities (23-26%) with glycerophospholipid cholesterol acyltransferase of Aeromonas spp. A recombinant PPP (rPPP) with a His tag at the C-terminus expressed in Escherichia coli and purified showed phospholipase activity. The rPPP also showed lecithin-dependent haemolytic activity against mammalian erythrocytes and direct haemolytic activity against fish erythrocytes. The culture supernatant of wild-type P. damselae ssp. piscicida showed phospholipase activity, while that of a PPP gene knockout mutant did not.
authors.affiliation: Laboratory of Genome Science, Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology, Tokyo, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Naka
authors.forename: I
authors.initials: I
authors.lastname: Hirono
authors.forename: T
authors.initials: T
authors.lastname: Aoki
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry; D004912:Erythrocytes / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005399:Fishes; D015870:Gene Expression / Q000502:physiology; D006461:Hemolysis / Q000502:physiology; D008322:Mammals; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D010740:Phospholipases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D010776:Photobacterium / Q000201:enzymology* / Q000235:genetics*; D010802:Phylogeny; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
chemical_list: D004269:DNA, Bacterial; D011994:Recombinant Proteins; D010740:Phospholipases