Cloning, purification and characterization of a functional anthracycline glycosyltransferase.
We have cloned the gene that encodes a novel glucosyl transferase (AraGT) involved in rhamnosylation of the polyketide antibiotic Aranciamycin in Streptomyces echinatus. AraGT comprises two domains characteristic of bacterial glycosyltranferases. AraGT was synthesized in E. coli as a decahistidinyl-tagged polypeptide. Purified AraGT is dimeric, displays a T(mapp) of 30 degrees C and can glycosylate the aglycone of an Aranciamycin derivative as shown by liquid chromatography and mass spectrometry. The availability of functional AraGT will allow the generation Aranciamycin-based combinatorial libraries.
Sianidis
Giorgos
G
Institute of Molecular Biology and Biotechnology, FORTH and Department of Biology, University of Crete, P.O. Box 1527, Iraklio-Crete 71110, Greece.
Wohlert
Sven-Eric
SE
Pozidis
Charalambos
C
Karamanou
Spyridoula
S
Luzhetskyy
Andriy
A
Vente
Andreas
A
Economou
Anastassios
A
D000595:Amino Acid Sequence; D018943:Anthracyclines / Q000378:metabolism*; D002853:Chromatography, Liquid; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D005964:Glucosyltransferases / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification*; D016695:Glycosyltransferases / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification*; D013058:Mass Spectrometry; D008954:Models, Biological; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics
D018943:Anthracyclines; D011994:Recombinant Proteins; C075925:aranciamycin; D016695:Glycosyltransferases; D005964:Glucosyltransferases; C515603:aranciamycin glucosyltransferase, Streptomyces echinatus