Incorporation of glucose analogs by GtfE and GtfD from the vancomycin biosynthetic pathway to generate variant glycopeptides.
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.
Losey
Heather C
HC
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA.
Jiang
Jiqing
J
Biggins
John B
JB
Oberthür
Markus
M
Ye
Xiang Yang
XY
Dong
Steven D
SD
Kahne
Daniel
D
Thorson
Jon S
JS
Walsh
Christopher T
CT
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
D000900:Anti-Bacterial Agents; D006020:Glycopeptides; D017334:Teicoplanin; D014640:Vancomycin; D005964:Glucosyltransferases; C113848:glucosyltransferase D; C430144:glucosyltransferase E; D005947:Glucose