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UBC Theses and Dissertations

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UBC Theses and Dissertations

Structural studies and bacteriophage degradation of Klebsiella capsular polysaccharides Lim, Andrew V. S.

Abstract

Seventy-seven serologically different strains of Klebsiella have been isolated. The capsular polysaccharides which these Gram-negative bacteria produce are antigenic. In order to understand the chemical basis of serological differentiation, the structural investigation of all 77 strains is taking place in this and other laboratories. To date, about sixty-five structures have been determined. The structure of the capsular antigen isolated from Klebsiella serotype K82 is presented here and was determined using the techniques of methylation, β-elimination, periodate oxidation-Smith hydrolysis and partial hydrolysis. Extensive use of n.m.r. spectroscopy (¹H and ¹³C) was used to establish the nature of the anomeric linkages of the polysaccharide and of derived poly- and oligo-saccharides obtained through degradative procedures. The K82 polysaccharide was found to comprise of tetrasaccharide repeating units of the 'three plus one' type: [diagram not included] The use of bacteriophage-borne glycanases as a technique in structural studies was demonstrated on Klebsiella K21 polysaccharide. Bacteriophage Φ21 was used to degrade K21 polysaccharide, resulting in a good yield of the single repeating unit. The structure determined for this bacteriophage-degraded product is in agreement with the previously known structure of K21 polysaccharide, which was determined solely by chemical methods.

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