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Updated: Jun 30, 2026

Identification and Characterization of Protein Glycosylation using Specific Endo- and Exoglycosidases
Published on: December 26, 2011
The in vitro characterization of polyene glycosyltransferases AmphDI and NysDI
Changsheng Zhang1, Rocco Moretti, Jiqing Jiang
1Laboratory for Biosynthetic Chemistry, Pharmaceutical Sciences Division, School of Pharmacy, UW-National Cooperative Drug Discovery Group, Program, University of Wisconsin-Madison, Madison, Wisconsin 53705-2222, USA.
Abstract:
The overproduction, purification, and in vitro characterization of the polyene glycosyltransferases (GTs) AmphDI and NysDI are reported. A novel nucleotidyltransferase mutant (RmlA Q83D) for the chemoenzymatic synthesis of unnatural GDP-sugar donors in conjunction with polyene GT-catalyzed sugar exchange/reverse reactions allowed the donor and acceptor specificities of these novel enzymes to be probed. The evaluation of polyene GT aglycon and GDP-sugar donor specificity revealed some tolerance to aglycon structural diversity, but stringent sugar specificity, and culminated in new polyene analogues in which L-gulose or D-mannose replace the native sugar D-mycosamine.
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