Biosynthetic gene cluster of the glycopeptide antibiotic teicoplanin: characterization of two glycosyltransferases and the key acyltransferase.
The gene cluster encoding biosynthesis of the clinically important glycopeptide antibiotic teicoplanin has been cloned from Actinoplanes teichomyceticus. Forty-nine putative open reading frames (ORFs) were identified within an 89 kbp genetic locus and assigned roles in teicoplanin biosynthesis, export, resistance, and regulation. Two ORFs, designated orfs 1 and 10*, showed significant homology to known glycosyltransferases. When heterologously expressed in Escherichia coli, these glycosyltransferases were shown to catalyze the transfer of UDP-(N-acetyl)-glucosamine onto, respectively, 3-chloro-beta-hydroxytyrosine-6 (3-Cl-6betaHty) and 4-hydroxyphenylglycine-4 (4Hpg) of the teicoplanin heptapeptide aglycone. The product of another ORF, orf11*, was demonstrated in vitro to transfer n-acetyl-, n-butyryl-, and n-octanoyl-groups from acyl-CoA donors either to a free UDP-aminosugar or to an aminosugar moiety in the teicoplanin pseudoaglycone, thus identifying Orf11* as the key acyltransferase in teicoplanin maturation. These findings should accelerate the combinatorial engineering of new and improved glycopeptide drugs.
Li
Tsung-Lin
TL
Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, United Kingdom.
Huang
Fanglu
F
Haydock
Stephen F
SF
Mironenko
Tatiana
T
Leadlay
Peter F
PF
Spencer
Jonathan B
JB
D000192:Actinomycetales / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D000217:Acyltransferases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000737:chemistry; D003001:Cloning, Molecular; D003500:Cyclization; D004351:Drug Resistance / Q000235:genetics; D005798:Genes, Bacterial; D016695:Glycosyltransferases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D006219:Halogens / Q000378:metabolism; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D015394:Molecular Structure; D005810:Multigene Family / Q000235:genetics; D017422:Sequence Analysis, DNA; D017334:Teicoplanin / Q000096:biosynthesis* / Q000737:chemistry
D000900:Anti-Bacterial Agents; D006219:Halogens; D017334:Teicoplanin; D000217:Acyltransferases; D016695:Glycosyltransferases