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not transfer GlcNAc and requires an unusual disaccharide. This
discovery complements prior GalNAc transfer and confirms a
predicted activity.18 Notably, WecA,43 the enzyme that would
generate PP-linked glycolipid substrates for PglB, is membrane-
associated and cannot be readily exploited in vitro. In vitro
biocatalytic installation of GlcNAc, shown here, also creates a
useful precursor sugar site for carbohydrate-processing enzyme-
mediated extension, as shown here (Scheme 2). The >90% in
vitro efficiencies shown here therefore make PglB a highly viable
synthetic biocatalyst for varied glycopolypeptides, coupled with
substrate accessibility and potential for further enzymatic
transformation.
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ASSOCIATED CONTENT
* Supporting Information
Full procedures. This material is available free of charge via the
■
S
(21) Jaenicke, L.; Siegmund, H. Biol. Chem. Hoppe-Seyler 1986, 367,
787.
AUTHOR INFORMATION
(22) Inoue, S.; Kaneko, T.; Takahashi, Y.; Miyamoto, O.; Sato, K. J.
Chem. Soc., Chem. Commun. 1987, 1036.
(23) Grassi, D.; Lippuner, V.; Aebi, M.; Brunner, J.; Vasella, A. J. Am.
Chem. Soc. 1997, 119, 10992.
(24) Chen, L.; Men, H.; Ha, S.; Ye, X.-Y.; Brunner, L.; Hu, Y.; Walker,
S. Biochemistry 2002, 41, 6824.
■
Corresponding Author
Author Contributions
§These authors contributed equally.
Notes
(25) Chang, Y.-F.; Liu, C.-Y.; Guo, C.-W.; Wang, Y.-C.; Fang, J.-M.;
Cheng, W.-C. J. Org. Chem. 2008, 73, 7197.
(26) Meng, F.-C.; Chen, K.-T.; Huang, L.-Y.; Shih, H.-W.; Chang, H.-
H.; Nien, F.-Y.; Liang, P.-H.; Cheng, T.-J. R.; Wong, C.-H.; Cheng, W.-
C. Org. Lett. 2011, 13, 5306.
The authors declare the following competing financial
interest(s): M.K. and A.F. are employees of Glycovaxyn. A
patent has been filed and will afford inventors royalties, if
licensed, in line with university guidelines.
(27) Hesek, D.; Lee, M.; Zajíce
Chem. Soc. 2012, 134, 13881.
̌
k, J.; Fisher, J. F.; Mobashery, S. J. Am.
ACKNOWLEDGMENTS
■
(28) Shih, H.-W.; Chang, Y.-F.; Li, W.-J.; Meng, F.-C.; Huang, C.-Y.;
Ma, C.; Cheng, T.-J. R.; Wong, C.-H.; Cheng, W.-C. Angew. Chem., Int.
Ed. 2012, 51, 10123.
(29) Lizak, C.; Gerber, S.; Numao, S.; Aebi, M.; Locher, K. P. Nature
2011, 474, 350.
(30) Jaenicke, L.; Siegmund, H.-U. Chem. Phys. Lipids 1989, 51, 159.
(31) Danilov, L. L.; Druzhinina, T. N.; Kalinchuk, N. A.; Maltsev, S. D.;
Shibaev, V. N. Chem. Phys. Lipids 1989, 51, 191.
(32) Bernardes, G. J. L.; Davis, B. G. unpublished results, University of
Oxford, 2007.
(33) Bernardes, G. J. L.; Kikkeri, R.; Maglinao, M.; Laurino, P.; Collot,
M.; Hong, S. Y.; Lepenies, B.; Seeberger, P. H. Org. Biomol. Chem. 2010,
8, 4987.
We thank the BBSRC (T.A.T.), Glycovaxyn (F.L., B.V., B.G.D.)
for funding, Drs P. Carranza, C. Bich for method development
and glycopeptide analysis, and Drs L. Gong, J. Wickens for
HRMS analysis. B.G.D. is a Royal Society Wolfson Merit
Research Award Recipient.
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