Complex polysaccharides on the surface of bacteria play a wide range of important roles in microbial survival and ecology. However, systems available to study their biosynthesis in cells are a major challenge. A majority of complex polysaccharides produced by bacteria are dependent on the C55 isoprenoid bactoprenyl diphosphate (BPP), which serves as a membrane localized anchor for the stepwise one sugar at-a-time assembly of these materials. Our aim is to develop new tools that exploit the dependence of these pathways on the polyisoprenoid to track the biosynthesis of complex polysaccharides in cells. A series of precursors to BPP have been synthesized and screened with undecaprenyl pyrophosphate synthase (UppS), which is responsible for the production of the isoprenoid in bacteria. In addition, a system has been developed as a cell-based method to determine whether these compounds can functionally replace important cellular precursors of BPP. We have found that new fluorescent polyprenol probes can be formed in cells that will be useful in the cellular analysis of any BPP dependent complex polysaccharide biosynthesis pathway, and may shed light on the critical cellular crosstalk between the many systems that are dependent on this critical molecule.<br/>

New tools for the elucidation of complex polysaccharide biosynthesis pathways

253rd National Meeting (2017)

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