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In vitro bacterial polysaccharide biosynthesis: defining the functions of Wzy and Wzz
Open accessAbstract
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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Citations
Cited by
Assembly of Bacterial Capsular Polysaccharides and Exopolysaccharides
Whitfield, C., Wear, S. S., & Sande, C. (2020) · Annual Review of Microbiology · doi:10.1146/annurev-micro-011420-075607 · Cited by 308
Enzymes in the Synthesis of Glycoconjugates
Schmaltz, R. M., Hanson, S. R., & Wong, C.-H. (2011) · Chemical Reviews · doi:10.1021/cr200113w · Cited by 283
Synthase-dependent exopolysaccharide secretion in Gram-negative bacteria
Whitney, J. C., & Howell, P. L. (2013) · Trends in Microbiology · doi:10.1016/j.tim.2012.10.001 · Cited by 266
Sharma, S., Erickson, K. M., & Troutman, J. M. (2017) · ACS Chemical Biology · doi:10.1021/acschembio.6b00931 · Cited by 37
Identification of an O‐antigen chain length regulator, WzzP, in Porphyromonas gingivalis
Shoji, M., Yukitake, H., Sato, K., Shibata, Y., Naito, M., Aduse‐Opoku, J., Abiko, Y., Curtis, M. A., & Nakayama, K. (2013) · MicrobiologyOpen · doi:10.1002/mbo3.84 · Cited by 30
Metadata
- Authors
- Woodward, R., Yi, W., Li, L., Zhao, G., Eguchi, H., Sridhar, P. R., Guo, H., Song, J. K., Motari, E., Cai, L., Kelleher, P., Liu, X., Han, W., Zhang, W., Ding, Y., Li, M., & Wang, P. G.
- Venue
- Nature Chemical Biology
- Published
- 2010-04-25
- Volume
- 6
- Issue
- 6
- Pages
- 418-423
- ISSN
- 1552-4450 (ISSN-L)
- 1552-4469
- OpenAlex
- W2055177969
- Cited by
- 175
- References
- 0
- FWCI
- 6.95
- Type
- journal article
- Language
- en
- Open access
- green
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- Woodward, R., Yi, W., Li, L., Zhao, G., Eguchi, H., Sridhar, P. R., Guo, H., Song, J. K., Motari, E., Cai, L., Kelleher, P., Liu, X., Han, W., Zhang, W., Ding, Y., Li, M., & Wang, P. G. (2010). In vitro bacterial polysaccharide biosynthesis: defining the functions of Wzy and Wzz. Nature Chemical Biology. https://doi.org/10.1038/nchembio.351
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