Bacteriophage-associated glycan hydrolases specific for Escherichia coli capsular serotype K12.
Four bacteriophages were identified, which carry glycan hydrolases specific for the Escherichia coli K12 capsular polysaccharide. All these glycanases catalyze the hydrolysis of the alpha-L-rhamnosyl-1,5-beta-3-deoxy-D-manno-2-octulosonic acid linkage as demonstrated with a special thiobarbituric acid assay procedure, which discriminates between the C5 substituted and unsubstituted 3-deoxy-D-manno-2-octulosonic acid (dOclA). This assay, together with gel filtration, 1H-NMR and 13C-NMR spectroscopy showed that depolymerization led to the dimer of the K12 repeating unit, (,5-beta-dOcl1Ap-2,3-alpha-LRhap-1,2-alpha LRhap-1,)2, as the primary degradation product. The phages (phi 12-W, phi 12-S, phi 82-W1, phi 82-W2) were tested for their ability to infect Escherichia coli strains Su65-42 (O4:K12:H-) and CDC63-57 [O139:K82(12):H1]. phi 12-W and phi 12-S, respectively, infected strain Su65-42 only, phi 82-W2 CDC63-57 only, and phi 82-W1 both bacterial strains. These distinct host specificities cannot be explained by differences in the action of the glycanases, which depolymerize the capsules of both strains.
Altmann
F
F
Institut für Chemie, Universität für Bodenkultur, Wien, Austria.
Christian
R
R
Czerny
T
T
Nimmich
W
W
März
L
L
D003090:Coliphages / Q000201:enzymology*; D004926:Escherichia coli / Q000145:classification / Q000201:enzymology / Q000276:immunology*; D006026:Glycoside Hydrolases / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D008970:Molecular Weight; D009844:Oligosaccharides / Q000302:isolation & purification; D011135:Polysaccharides, Bacterial / Q000378:metabolism*; D012703:Serotyping; D013379:Substrate Specificity
D009844:Oligosaccharides; D011135:Polysaccharides, Bacterial; D006026:Glycoside Hydrolases