Two different O-polysaccharides from Escherichia coli O86 are produced by different polymerization of the same O-repeating unit

Yi, Wen; Bystrický, Peter; Yao, Qian; Guo, Hongjie; Zhu, Lizhi; Li, Hanfen; Shen, Jie; Li, Mei; Ganguly, Soumya; Bush, C. Allen; Wang, Peng George

doi:10.1016/j.carres.2005.11.001

Cited by 26 publications

(21 citation statements)

References 34 publications

(39 reference statements)

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“…According to methylation data, typical signal at 2.01 ppm was to be ascribed to the N-acetyl group of GlcpNAc (Nordmark et al, 2005). H-6 signals at 1.33 and 1.19 ppm can be explained by the presence of the methyl groups of Rhap and/or Fucp units (Karlsson et al, 1997;Yi et al, 2006). Moreover, the amino acid analysis indicated that it contained 16 types of amino acid.…”

Section: Discussionmentioning

confidence: 92%

Effects of Gekko sulfated polysaccharide–protein complex on human hepatoma SMMC-7721 cells: Inhibition of proliferation and migration

Chen

Yao

Zhang

et al. 2010

Journal of Ethnopharmacology

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Section: Discussionmentioning

confidence: 92%

Effects of Gekko sulfated polysaccharide–protein complex on human hepatoma SMMC-7721 cells: Inhibition of proliferation and migration

Chen

Yao

Zhang

et al. 2010

Journal of Ethnopharmacology

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“…To investigate this hypothesis in vitro , we introduced purified Wzz proteins obtained from two different E. coli O86 strains (B7 and H2) into the polymerization system. O-polysaccharides from these two strains share the same branched pentasaccharide repeating unit but differ in the anomeric configuration of the linkage25. Furthermore, although Wzz proteins from these two strains share 90% amino acid identity, the LPS modality of the two strains is markedly different.…”

Section: Resultsmentioning

confidence: 99%

“…The O-polysaccharide-PP-Und generated by in vitro Wzy-Wzz reconstitution ( a3 and a4 ) exhibits a modality similar to that of LPS regulated by Wzz B7 and Wzz H2 ( b1 and b3 ), respectively. It should also be noted that the O-polysaccharides from these two strains share the same branched pentasaccharide repeating unit but differ in the anomeric configuration of the linkage25.…”

Section: Figurementioning

confidence: 99%

In vitro bacterial polysaccharide biosynthesis: defining the functions of Wzy and Wzz

et al. 2010

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Polysaccharides constitute a major component of bacterial cell surfaces and play critical roles in bacteria/host interactions. The biosynthesis of such molecules, however, has mainly been characterized through in vivo genetic studies, thus precluding discernment of the details of this pathway. Accordingly, we present a chemical approach which enabled reconstitution of the E. coli O-polysaccharide biosynthetic pathway in vitro. Starting with chemically prepared N-Acetyl-D-galactosamine-diphospho-undecaprenyl, the E. coli O86 oligosaccharide repeating unit was assembled via sequential enzymatic glycosylation. Successful expression of the putative polymerase Wzy via a chaperone co-expression system then allowed demonstration of polymerization in vitro using this substrate. Analysis of additional substrates revealed a defined mode of recognition for Wzy towards the lipid moiety. Specific polysaccharide chain length modality was furthermore demonstrated to result from the action of Wzz. Collectively, polysaccharide biosynthesis was chemically reconstituted in vitro, providing a well-defined system for further underpinning molecular details of this biosynthetic pathway.

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“…Fucose-containing E. coli O86 O-antigen (lipopolysaccharide, LPS), whose structure was solved by our group (9), was used as a model system. We chose to introduce fucose analogs into this polysaccharide to demonstrate our strategy.…”

Section: Resultsmentioning

confidence: 99%

Remodeling bacterial polysaccharides by metabolic pathway engineering

Liu

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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Introducing structural modifications into biomolecules represents a powerful approach to dissect their functions and roles in biological processes. Bacterial polysaccharides, despite their rich structural information and essential roles in bacterium-host interactions and bacterial virulence, have largely been unexplored for in vivo structural modifications. In this study, we demonstrate the incorporation of a panel of monosaccharide analogs into bacterial polysaccharides in a highly homogenous manner via metabolic engineering of a promiscuous sugar nucleotide biosynthetic pathway. In addition, the bioorthorgonal functional groups metabolically incorporated were exploited for cell surface labeling using in vitro selective chemical ligation reactions. In summary, our study presents a general, facile and effective approach for in vivo generation of novel tailor-made bacterial polysaccharides.chemical remodeling ͉ metabolic engineering ͉ sugar nucleotide biosynthesis ͉ fucose

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Two different O-polysaccharides from Escherichia coli O86 are produced by different polymerization of the same O-repeating unit

Cited by 26 publications

References 34 publications

Effects of Gekko sulfated polysaccharide–protein complex on human hepatoma SMMC-7721 cells: Inhibition of proliferation and migration

Effects of Gekko sulfated polysaccharide–protein complex on human hepatoma SMMC-7721 cells: Inhibition of proliferation and migration

In vitro bacterial polysaccharide biosynthesis: defining the functions of Wzy and Wzz

Remodeling bacterial polysaccharides by metabolic pathway engineering

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