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Identification ofStreptomyces violaceoruber Tü22 genes involved in the biosynthesis of granaticin

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Abstract

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.

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Author notes

  1. Andreas Bechthold

    Present address: Institut für Pharmazeutische Biologie, Eberhard-Karls-Universität Tübingen, 72076, Tübingen, Germany

  2. Jae Kyung Sohng

    Present address: Department of Chemistry, Sunmoon University, Chunahm, Sambyongdong 381-7, 330-150, Chungnam, Korea

  3. Xin Chu

    Present address: Department of Microbiology, North Dakota State University, 241 E. University Village, 58102, Fargo, ND, USA

  4. Todd M. Smith

    Present address: Department of Molecular Biotechnology FJ-20, University of Washnigton, 98195, Seattle, WA, USA

Authors and Affiliations

  1. Department of Chemistry, University of Washington, 98195, Seattle, WA, USA

    Andreas Bechthold, Jae Kyung Sohng, Xin Chu & Heinz G. Floss

  2. Department of Medicinal Chemistry, University of Washington, 98195, Seattle, WA, USA

    Todd M. Smith & Heinz G. Floss

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  1. Andreas Bechthold
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  2. Jae Kyung Sohng
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  5. Heinz G. Floss
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Communicated by W. Goebel

Dedicated to Professor Satoshi Ōmura, a pioneer in the field of antibiotics, on the occasion of his 60th birthday

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Bechthold, A., Sohng, J.K., Smith, T.M. et al. Identification ofStreptomyces violaceoruber Tü22 genes involved in the biosynthesis of granaticin. Molec. Gen. Genet. 248, 610–620 (1995). https://doi.org/10.1007/BF02423457

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  • DOI: https://doi.org/10.1007/BF02423457

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