title: Incorporation of glucose analogs by GtfE and GtfD from the vancomycin biosynthetic pathway to generate variant glycopeptides.
abstract: Analogs of the glycopeptide antibiotics vancomycin and teicoplanin with alterations in one or both sugar moieties of the disaccharide have been prepared by tandem action of the vancomycin pathway glycosyltransferases GtfE and GtfD. All four regioisomers (2-, 3-, 4-, 6-) of TDP-deoxyglucoses and UDP/TDP-aminoglucoses were prepared, predominantly by action of D-glucopyranosyl-1-phosphate thymidylyltransferase, E(p). GtfE transferred the deoxyglucoses or aminoglucoses onto the 4-OH of 4-hydroxyphenylglycine of both the vancomycin and teicoplanin aglycone scaffolds. Kinetic analysis indicated the 2-, 3-, 4-, and 6-amino-glucoses were transferred by GtfE with only a 4- to 30-fold drop in k(cat) and no effect on K(m) compared to the native substrate, UDP/TDP-glucose, suggesting preparative utility. The next enzyme, GtfD, could utilize the variant glucosyl-peptides as substrates for transfer of L-4-epi-vancosamine. The aminosugar moieties in these variant glycopeptides introduce sites for acylation or reductive alkylation.
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Heather C
authors.initials: HC
authors.lastname: Losey
authors.forename: Jiqing
authors.initials: J
authors.lastname: Jiang
authors.forename: John B
authors.initials: JB
authors.lastname: Biggins
authors.forename: Markus
authors.initials: M
authors.lastname: Oberthür
authors.forename: Xiang Yang
authors.initials: XY
authors.lastname: Ye
authors.forename: Steven D
authors.initials: SD
authors.lastname: Dong
authors.forename: Daniel
authors.initials: D
authors.lastname: Kahne
authors.forename: Jon S
authors.initials: JS
authors.lastname: Thorson
authors.forename: Christopher T
authors.initials: CT
authors.lastname: Walsh
mesh_terms: D000900:Anti-Bacterial Agents / Q000096:biosynthesis / Q000737:chemistry; D020650:Combinatorial Chemistry Techniques; D005947:Glucose / Q000031:analogs & derivatives* / Q000378:metabolism; D005964:Glucosyltransferases / Q000378:metabolism*; D006020:Glycopeptides / Q000096:biosynthesis* / Q000737:chemistry; D007700:Kinetics; D015394:Molecular Structure; D013379:Substrate Specificity; D017334:Teicoplanin / Q000031:analogs & derivatives / Q000096:biosynthesis; D014640:Vancomycin / Q000031:analogs & derivatives* / Q000096:biosynthesis
chemical_list: D000900:Anti-Bacterial Agents; D006020:Glycopeptides; D017334:Teicoplanin; D014640:Vancomycin; D005964:Glucosyltransferases; C113848:glucosyltransferase D; C430144:glucosyltransferase E; D005947:Glucose