Enzymatic action of coliphage omega8 and its possible role in infection.
The receptor of coliphage omega8 is the O-specific mannan of Escherichia coli O8 in which the trisaccharide alpha-mannosyl-1,2-alpha-mannosyl-1,2-mannose is joined through alpha-mannosyl-1,3-linkages. Coliphage omega8 produces an endo-alpha-1,3-mannosidase which destroys the receptor, liberating a series of oligosaccharides (repeating trisaccharide and multiples). The enzyme is an integral part of the phage particles and also occurs in a free form in the lysates. Phage particles hydrolyze alpha-1,3-mannosyl linkages in the lipopolysaccharide, the polysaccharide (mannan) moiety, and higher oligosaccharides with an efficiency decreasing in this order. No transmannosylation could be detected. Phage particles also degrade the receptor mannan on whole bacteria, as determined with 14C-labeled E. coli O8. The values of Km and Vmax were determined with omega8 particles and free enzymes using native lipopolysaccharide and its triethylammonium salt. The latter, which was obtained after electrodialysis, has a micellar weight of 2.5 X 10(5), whereas the native lipopolysaccharide forms supermicelles with micellar weights of several millions. With coliphage omega8 as enzyme and supermicellar lipopolysaccharide as substrate Km=5 X 10(-8) M was obtained. This, together with the fact that omega8 attaches irreversibly to E. coli O8, was used in proposing a hypothesis for the possible role of the enzyme in the first steps of infection with coliphage omega8.
D000327:Adsorption; D001665:Binding Sites; D003090:Coliphages / Q000201:enzymology*; D004186:Disaccharidases / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008070:Lipopolysaccharides / Q000378:metabolism; D008351:Mannans / Q000378:metabolism; D008361:Mannosidases / Q000378:metabolism*; D011135:Polysaccharides, Bacterial / Q000378:metabolism
D008070:Lipopolysaccharides; D008351:Mannans; D011135:Polysaccharides, Bacterial; D004186:Disaccharidases; D008361:Mannosidases