1887

Abstract

-Rhamnose (-Rha) is a component of the lipopolysaccharide (LPS) core, several O antigen polysaccharides, and the cell surface surfactant rhamnolipid of . In this study, four contiguous genes () responsible for the synthesis of dTDP--Rha in have been cloned and characterized. Non-polar chromosomal mutants were generated in strains PAO1 (serotype O5) and PAK (serotype O6) and LPS extracted from the mutants was analysed by SDS-PAGE and Western immunoblotting. mutants of both serotype O5 and serotype O6 synthesized a truncated core region which was unable to act as an attachment point for either A-band or B-band O antigen. A PAO1 double mutant (deficient in biosynthesis of both -Rha and -Rha) was constructed to facilitate structural analysis of the mutant core region. This strain has an incomplete core oligosaccharide region and does not produce A-band O antigen. These results provide the genetic and structural evidence that -Rha is the receptor on the LPS core for the attachment of O polysaccharides. This is the first report of a genetically defined mutation that affects the synthesis of a single sugar in the core oligosaccharide region of LPS, and provides further insight into the mechanisms of LPS biosynthesis and assembly in this bacterium.

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2000-11-01
2019-12-11
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