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Abstract

Polysialic acids occur as capsular polysaccharides of several pathogenic bacteria. An understanding of how polysialyltransferase functions in the synthesis of polysialic acid will require enzyme purification and characterization in concert with genetic analysis. A rapid filter assay has been developed for bacterial polysialyltransferase suitable for enzyme purification. The filter assay and the currently used paper chromatography methods are equivalent in parallel experiments. The Escherichia coli K92 polysialyltransferase exhibited the same pH and temperature optima, Mg2+ dependence and acceptor specificity in both assays. [14C]Sialic acid bound in filter assays correlates with polymer formed by gel filtration. Specificity may be increased by the addition of exogenous acceptors.

Sialyltransferase, Polysialic acid biosynthesis, Enzyme assay, Escherichia coli

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© 1995 Federation of European Microbiological Societies
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