Cloning and characterization of a glycosyltransferase gene involved in the biosynthesis of anthracycline antibiotic β-rhodomycin from Streptomyces violaceus

A glycosyltransferase gene, rhoG, involved in the biosynthesis of the anthracycline antibiotic β-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 g...

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Bibliographic Details
Published in:FEMS microbiology letters 2002-01, Vol.206 (2), p.163-168
Main Authors: Miyamoto, Yuji, Johdo, Osamu, Nagamatsu, Yasunori, Yoshimoto, Akihiro
Format: Article
Language:English
Subjects:
Anthracycline
anthracycline antibiotics
Anthracyclines - metabolism
Antibiotics, Antineoplastic - metabolism
b-Rhodomycin
Bacterial Proteins
Bacteriology
Biological and medical sciences
Biosynthesis
Cloning, Molecular
Fundamental and applied biological sciences. Psychology
Genes, Bacterial
Genetic Complementation Test
Genetics
Glycosylation
Glycosyltransferase
Glycosyltransferases - genetics
Microbiology
Mutagenicity Tests
rhoG gene
Sequence Analysis, DNA
Streptomyces - enzymology
Streptomyces - genetics
Streptomyces gene
Streptomyces violaceus
β-Rhodomycin
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Summary:A glycosyltransferase gene, rhoG, involved in the biosynthesis of the anthracycline antibiotic β-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 β-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 β-rhodomycin productivity, along with the accumulation of a non-glycosyl intermediate ϵ-rhodomycinone. In addition, the complementation test demonstrated that rhoG restored β-rhodomycin production in this gene disruptant. These results indicated that rhoG is the glycosyltransferase gene responsible for the glycosylation of ϵ-rhodomycinone in β-rhodomycin biosynthesis.
ISSN:0378-1097
1574-6968
DOI:10.1016/S0378-1097(01)00531-6