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Cloning and characterization of Photobacterium damselae ssp. piscicida phospholipase: an enzyme that shows haemolytic activity
Author(s) -
Naka H,
Hirono I,
Aoki T
Publication year - 2007
Publication title -
journal of fish diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.819
H-Index - 85
eISSN - 1365-2761
pISSN - 0140-7775
DOI - 10.1111/j.1365-2761.2007.00861.x
Subject(s) - biology , biochemistry , phospholipase a , phospholipase , photobacterium , open reading frame , enzyme , molecular cloning , vibrio , phospholipase a1 , microbiology and biotechnology , peptide sequence , gene , phospholipase a2 , bacteria , genetics
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.