Identification and characterisation of hyaluronate lyase from Streptococcus suis.
Hyaluronate lyase, which catalyses the degradation of hyaluronic acid (HA), has been described from several pathogenic streptococcal species. We describe, for the first time, identification and purification of hyaluronate lyase from the zoonotic pig pathogen Streptococcus suis. We have cloned the hyaluronate lyase gene from S. suis and used it to generate an allelic replacement knock-out mutant of S. suis serotype 7 that can no longer biosynthesise the enzyme. Interestingly, a limited strain survey indicates that hyaluronate lyase activity is not present in all disease isolates of S. suis. Polyclonal anti-hyaluronate lyase anti-serum raised against our recombinant hyaluronate lyase has been used in Western blots, showing that hyaluronate lyase activity is always associated with the presence of protein of the expected size, whereas lack of hyaluronate lyase activity is due to truncation or absence of the enzyme. We show that hyaluronate lyase activity is required for S. suis to use HA polymer as a carbon source and that supplying exogenous recombinant hyaluronate lyase to all S. suis strains tested allowed fermentation of the resultant HA breakdown products.
Allen
Andrew G
AG
Department of Clinical Veterinary Medicine, Centre for Veterinary Science, University of Cambridge, Madingley Road, Cambridge CB3 0ES, UK.
Lindsay
Heather
H
Seilly
David
D
Bolitho
Steven
S
Peters
Sarah E
SE
Maskell
Duncan J
DJ
D015153:Blotting, Western; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification; D017353:Gene Deletion; D005798:Genes, Bacterial; D006820:Hyaluronic Acid / Q000378:metabolism; D008969:Molecular Sequence Data; D011133:Polysaccharide-Lyases / Q000032:analysis / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000276:immunology; D017422:Sequence Analysis, DNA; D017011:Streptococcus suis / Q000201:enzymology* / Q000302:isolation & purification; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism
D004269:DNA, Bacterial; D011994:Recombinant Proteins; D037521:Virulence Factors; D006820:Hyaluronic Acid; D011133:Polysaccharide-Lyases; C100002:hyaluronate lyase