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O-antigen polymerase adopts a distributive mechanism for lipopolysaccharide biosynthesis

Applied Microbiology and Biotechnology volume 98, pages 4075–4081 (2014)Cite this article

An Erratum to this article was published on 02 April 2014

Abstract

Bacterial lipopolysaccharide (LPS) is an essential cell envelope component for gram-negative bacteria. As the most variable region of LPS, O antigens serve as important virulence determinants for many bacteria and represent a promising carbohydrate source for glycoconjugate vaccines. In the Wzy-dependent O-antigen biosynthetic pathway, the integral membrane protein Wzy was shown to be the sole enzyme responsible for polymerization of O-repeat unit. Its catalytic mechanism, however, remains elusive. Herein, Wzy was successfully overexpressed in Escherichia coli with an N-terminal His10-tag. Blue native polyacrylamide gel electrophoresis (BN-PAGE) revealed that the Wzy protein exists in its native confirmation as a dimer. Subsequently, we chemo-enzymatically synthesized the substrates of Wzy, the lipid-PP-linked repeat units. Together with an optimized O-antigen visualization method, we monitored the production of reaction intermediates at varying times. We present here our result as the first biochemical evidence that Wzy functions in a distributive manner.

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Acknowledgement

P.G.W. acknowledges the NIH R01 GM085267 for financial support.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Center for Diagnostics and Therapeutics and Department of Chemistry, Georgia State University, Atlanta, GA, 30303, USA

    Guohui Zhao, Baolin Wu, Lei Li & Peng George Wang

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  1. Guohui Zhao
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  2. Baolin Wu
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  3. Lei Li
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  4. Peng George Wang
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Correspondence to Peng George Wang.

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Zhao, G., Wu, B., Li, L. et al. O-antigen polymerase adopts a distributive mechanism for lipopolysaccharide biosynthesis. Appl Microbiol Biotechnol 98, 4075–4081 (2014). https://doi.org/10.1007/s00253-014-5552-7

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  • DOI: https://doi.org/10.1007/s00253-014-5552-7

Keywords

  • Wzy
  • Integral membrane protein
  • Overexpression
  • Distributive mechanism