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Identification and organization of genes for diutan polysaccharide synthesis from Sphingomonas sp. ATCC 53159

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Journal of Industrial Microbiology & Biotechnology

Abstract

A cluster of genes for diutan polysaccharide synthesis was isolated from a library of Sphingomonas sp. ATCC 53159 genomic DNA by complementation of glucosyl-isoprenylphosphate transferase-deficient mutants of Sphingomonas elodea ATCC 31461 (producing gellan) and Xanthomonas campestris (producing xanthan). The synthesis of polysaccharide in these strains shares a common first step, transfer of glucose-1-phosphate from UDP-glucose to the isoprenylphosphate lipid. The cluster of 24 genes was compared to genes for biosynthesis of gellan, and S-88 sphingan from Sphingomonas sp. ATCC 31554. Diutan, gellan and S-88 sphingan have a common four-sugar backbone but different side chains, one rhamnose for S-88 sphingan, a two-rhamnose side chain for diutan and no side chain for gellan. The genes for biosynthesis of diutan, gellan and S-88 sphingan were similar in general organization but differed in location of some genes, in particular, dpsG (putative polymerase), dpsR (putative lyase) and dpsS (putative repeat unit transporter). An unidentified reading frame urf31, present in the gene clusters for diutan and S-88 sphingan but not gellan, had similarity to glycosyl transferase group 2 proteins, and was detrimental when cloned in Sphingomonas elodea producing gellan that lacks a side chain, but not in Sphingomonas ATCC 31554 producing S-88 sphingan with a rhamnose side chain. Gene urf31 could possibly encode a side-chain rhamnosyl transferase. Another gene urf31.4 was unique to the diutan gene cluster. A plasmid containing 20 of the 24 genes resulted in a slight increase in the amount of diutan produced, but a significant increase in the rheological properties of diutan.

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Acknowledgments

We thank Luis Ielpi for his careful reading of this manuscript and for supplying some of the plasmids used in this study. We thank Anca Segall, Judith Zyskind and Luis Ielpi for helpful discussions. We appreciate the technical assistance of German Berrellez and Peter Mirrasoul for fermentation analysis, Steve Matzke and Dan Burgum for rheological analysis, and M.R. Abouzari for composition analysis.

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  1. Russell J. Coleman

    Present address: The Dow Chemical Company, 5501 Oberlin Drive, San Diego, CA, 92121, USA

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  1. Molecular Biology, CP Kelco, 8225 Aero Drive, San Diego, CA, 92123, USA

    Russell J. Coleman, Yamini N. Patel & Nancy E. Harding

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  1. Russell J. Coleman
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  2. Yamini N. Patel
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  3. Nancy E. Harding
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Correspondence to Nancy E. Harding.

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Coleman, R.J., Patel, Y.N. & Harding, N.E. Identification and organization of genes for diutan polysaccharide synthesis from Sphingomonas sp. ATCC 53159. J Ind Microbiol Biotechnol 35, 263–274 (2008). https://doi.org/10.1007/s10295-008-0303-3

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