A Systematic Investigation of the Synthetic Utility of Glycopeptide Glycosyltransferases

• Published in: Journal of the American Chemical Society Vol. 127; no. 30; pp. 10747 - 10752
• Main Authors: Oberthür, Markus, Leimkuhler, Catherine, Kruger, Ryan G, Lu, Wei, Walsh, Christopher T, Kahne, Daniel
• Format: Journal Article
• Language: English
• Published: WASHINGTON American Chemical Society 03.08.2005; Amer Chemical Soc
• Subjects: Anti-Bacterial Agents - biosynthesis; Anti-Bacterial Agents - chemical synthesis; Antibacterial agents; Antibiotics. Antiinfectious agents. Antiparasitic agents; Biological and medical sciences; Chemistry; Chemistry, Multidisciplinary; Exact sciences and technology; Glycosylation; Glycosyltransferases - chemistry; Glycosyltransferases - isolation & purification; Glycosyltransferases - metabolism; Hexosamines - chemistry; Hexosamines - metabolism; Isoenzymes; Kinetics; Medical sciences; Organic chemistry; Peptides; Pharmacology. Drug treatments; Physical Sciences; Preparations and properties; Science & Technology; Substrate Specificity; Vancomycin - analogs & derivatives; Vancomycin - chemical synthesis; Vancomycin - chemistry; Vancomycin - metabolism; VANCOMYCIN ANALOGS; SUBSTRATE-SPECIFICITY; SUGAR NUCLEOTIDES; ANTIBIOTICS; TANDEM ACTION; STAPHYLOCOCCUS-AUREUS; BIOSYNTHETIC PATHWAYS; GLYCORANDOMIZATION; GLYCOSYLATION; DERIVATIVES; Enzyme; Transferases; Glycosyltransferases; Biosynthesis; Glycosylation; In vitro; Biological activity; Antibiotic; Transfer reaction; Glycopeptide; Peptide synthesis; Antibacterial agent; Enzymatic reaction
• ISSN: 0002-7863; 1520-5126
• DOI: 10.1021/ja052945s

Glycosyltransferases involved in the biosynthesis of bacterial secondary metabolites may be useful for the generation of sugar-modified analogues of bioactive natural products. Some glycosyltransferases have relaxed substrate specificity, and it has been assumed that promiscuity is a feature of the class. As part of a program to explore the synthetic utility of these enzymes, we have analyzed the substrate selectivity of glycosyltransferases that attach similar 2-deoxy-l-sugars to glycopeptide aglycons of the vancomycin-type, using purified enzymes and chemically synthesized TDP β-2-deoxy-l-sugar analogues. We show that while some of these glycopeptide glycosyltransferases are promiscuous, others tolerate only minor modifications in the substrates they will handle. For example, the glycosyltransferases GtfC and GtfD, which transfer 4-epi-l-vancosamine and l-vancosamine to C-2 of the glucose unit of vancomycin pseudoaglycon and chloroorienticin B, respectively, show moderately relaxed donor substrate specificities for the glycosylation of their natural aglycons. In contrast, GtfA, a transferase attaching 4-epi-l-vancosamine to a benzylic position, only utilizes donors that are closely related to its natural TDP sugar substrate. Our data also show that the spectrum of donors utilized by a given enzyme can depend on whether the natural acceptor or an analogue is used, and that GtfD is the most versatile enzyme for the synthesis of vancomycin analogues.