Cloning and characterization of a glycosyltransferase gene involved in the biosynthesis of anthracycline antibiotic beta-rhodomycin from Streptomyces violaceus.
A glycosyltransferase gene, rhoG, involved in the biosynthesis of the anthracycline antibiotic beta-rhodomycin was isolated as a 4.1-kb DNA fragment containing rhoG and its flanking region from Streptomyces violaceus by degenerate and inverse PCR. Sequencing analysis showed that rhoG was located in a gene cluster involved in the biosynthesis of the constitutive deoxysugar of beta-rhodomycin. The function of rhoG was verified by gene disruption, which was generated by replacing the internal 0.9-kb region of S. violaceus chromosome with a fragment including the SacI-blunted region. The rhoG disruption resulted in complete loss of beta-rhodomycin productivity, along with the accumulation of a non-glycosyl intermediate epsilon-rhodomycinone. In addition, the complementation test demonstrated that rhoG restored beta-rhodomycin production in this gene disruptant. These results indicated that rhoG is the glycosyltransferase gene responsible for the glycosylation of epsilon-rhodomycinone in beta-rhodomycin biosynthesis.
Miyamoto
Yuji
Y
Department of Applied Biochemistry, Faculty of Applied Biological Sciences, Hiroshima University, Japan. yujim@hiroshima-u.ac.jp
Johdo
Osamu
O
Nagamatsu
Yasunori
Y
Yoshimoto
Akihiro
A
D018943:Anthracyclines / Q000378:metabolism*; D000903:Antibiotics, Antineoplastic / Q000378:metabolism*; D001426:Bacterial Proteins; D003001:Cloning, Molecular; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D006031:Glycosylation; D016695:Glycosyltransferases / Q000235:genetics*; D009152:Mutagenicity Tests; D017422:Sequence Analysis, DNA; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics*
D018943:Anthracyclines; D000903:Antibiotics, Antineoplastic; D001426:Bacterial Proteins; C004977:rhodomycin; D016695:Glycosyltransferases; C449814:RhoG protein, Streptomyces violaceus