Identification ofStreptomyces violaceoruber Tü22 genes involved in the biosynthesis of granaticin
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A 50 kb region of DNA fromStreptomyces violaceoruber Tü22, containing genes encoding proteins involved in the biosynthesis of granaticin, was isolated. The DNA sequence of a 7.3 kb fragment from this region, located approximately 10 kb from the genes that encode the polyketide synthetase responsible for formation of the benzoisochromane quinone skeleton, revealed five open reading frames (ORF1-ORF5). The deduced amino acid sequence of GraE, encoded by ORF2, shows 60.8% identity (75.2% similarity) to a dTDP-glucose dehydratase (StrE) fromStreptomyces griseus. Cultures ofEscherichia coli containing plasmids with ORF2, on a 2.1 kbBamHI fragment, were able to catalyze the formation of dTDP-4-keto-6-deoxy-d-glucose from dTDP-glucose at 5 times the rate of control cultures, confirming that ORF2 encodes a dTDP-glucose dehydratase. The amino acid sequence encoded by ORF3 (GraD) is 51.4% identical (69.9% similar) to that of StrD, a dTDP-glucose synthase fromStreptomyces griseus. The amino acid sequence encoded by ORF4 shares similarities with proteins that confer resistance to tetracycline and methylenomycin, and is suggested to be involved in transporting granaticin out of the cells by an active efflux mechanism.
KeywordsAmino Acid Sequence Tetracycline Quinone Deduce Amino Acid Sequence Control Culture
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- Calcutt MJ, Schmidt FJ (1991) Bleomycin biosynthesis: characterization of the resistance region of the producer streptomycete. International Symposium on Biology of Actinomycetes, University of Wisconsin, Madison wis USAGoogle Scholar
- He X-G, Chang C-C, C-J. C, Vederas JC, McInnes AG, Walter JA, Floss HG (1986) Further studies on the biosynthesis of granaticin. Z Naturforsch 41c:215–221Google Scholar
- Hopwood DA, Bibb MJ, Chater KF, Kieser T, Bruton C, Kieser HM, Lydiate DJ, Smith CP, Ward JM (1985) Genetic manipulation ofStreptomyces: a laboratory manual. John Innes Foundation, Norwich, UKGoogle Scholar
- Hui-Zahn Z, Schmidt H, Piepersberg W (1992) Molecular cloning and characterization of two genes fromStreptomyces lincolnensis 78-11. Mol Microbiol 6:3147–3157Google Scholar
- Matsudaira PT (1993). A practical guide to protein and peptide purification for microsequencing. Academic Press, San DiegoGoogle Scholar
- Normark S, Bergström S, Edlund T, Grundström T, Jaurin B, Lindberg FP, Olsson O (1983) Overlapping genes. Annu Rev Genet 58:229–241Google Scholar
- Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New YorkGoogle Scholar
- Stockmann M, Piepersberg W (1992) Gene probes for the detection of 6-deoxyhexose metabolism in secondary metabolite-producingStreptomycetes. FEMS Microbiol Lett 90:185–190Google Scholar