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Cloning and nucleotide sequence of fosfomycin biosynthetic genes of Streptomyces wedmorensis

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Abstract

The biosynthetic pathway for production of the antibiotic fosfomycin by Streptomyces wedmorensis consists of four steps including the formation of a C-P bond and an epoxide. Fosfomycin production genes were cloned from genomic DNA using S. wedmorensis mutants blocked at different steps of the biosynthetic pathway. Four genes corresponding to each of the biosynthetic steps were found to be clustered in a DNA fragment of about 5 kb. Nucleotide sequencing of a large fragment revealed the presence of ten open reading frames, including the four biosynthetic genes and six genes with unknown functions.

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References

  • Hammerschmidt F (1991) Biosynthesis of natural products with a C-P bond. Part 8: On the origin of the oxirane oxygen atom of fosfomycin in Streptomyces fradiae. J Chem Soc Perkin Trans 1:1993–1996

    Google Scholar 

  • Hammerschmidt F, Bovermann G, Bayer K (1990) Das Oxiran-Sauerstoff-Atom des fosfomycins entstammt nicht dem luft-sauerstoff. Liebigs Ann Chem 1055–1061

  • Hashimoto T, Yamada Y (1987) Purification and characterization of hyoscyamine 6β-hydroxylase from root cultures of Hyoscyamus niger L. Eur J Biochem 164:277–285

    Google Scholar 

  • Hashimoto T, Yamada Y (1992) Biosynthesis of scopolamine and an application for genetic engineering of medical plant. Plant Tissue Culture and Gene Manipulation for Breeding and Formation of Phytochemicals, pp 255–259

  • Hashimoto T, Hayashi A, Amano Y, Kohno J, Iwanari H, Usuda S, Yamada Y (1991) Hyoscyamine 6β-hydroxylase, an enzyme involved in tropane alkaloid biosynthesis, is localized at the pericycle of the root. J Biol Chem 266:4648–4653

    Google Scholar 

  • Hendlin D, Stapley EO, Jackson M, Wallick H, Miller AK, Wolf FJ, Miller TW, Chaiet L, Kahan FM, Foltz EL, Woodruff HB, Mata JM, Hernandez S, Mochales S (1969) Phosphonomycin, a new antibiotic produced by strains of Streptomyces. Science 166:122–123

    Google Scholar 

  • Hidaka T, Mori M, Imai S, Hara O, Nagaoka K, Seto H (1989) Studies on the biosynthesis of bialaphos (SF-1293). 9. Biochemical mechanism of C-P bond formation in bialaphos: discovery of phosphoenolpyruvate phosphomutase which catalyzes the formation of phosphonopyruvate from phosphoenolpyruvate. J Antibiot 42:491–494

    Google Scholar 

  • Hidaka T, Iwaskura H, Imai S, Seto H (1992a) Studies on the biosynthesis of fosfomycin. 3. Detection of phosphoenolpyruvate phosphomutase activity in a fosfomycin high-producing strain of Streptomyces wedmorensis and characterization of its blocked mutant NP-7. J Antibiot 45:1008–1010

    Google Scholar 

  • Hidaka T, Hidaka M, Seto H (1992b) Studies on the biosynthesis of bialaphos (SF-1293). 14. Nucleotide sequence of phosphoenolpyruvate phosphomutase gene isolated from a bialaphos producing organism, Streptomyces hygroscopicus, and its expression in Streptomyces lividans. J Antibiot 45:1977–1980

    Google Scholar 

  • Hidaka T, Hidaka M, Uozumi T, Seto H (1992c) Nucleotide sequence of carboxyphosphonoenolpyruvate phosphomutase gene isolated from a bialaphos producing organism, Streptomyces hygroscopicus, and its expression in Streptomyces lividans. Mol Gen Genet 233:476–478

    Google Scholar 

  • Hidaka T, Hidaka M, Seto H (1995) Sequence of a P-methyltransferase-encoding gene isolated from a bialaphos-producing Streptomyces hygroscopicus. Gene, in press

  • Hopwood DA, Bibb MJ, Charter KF, Kieser T, Bruton CJ, Kieser HM, Lydiate DJ, Smith CP, Ward JM, Schrempf H (1985) Genetic manipulation of Streptomyces. A laboratory manual. The John Innes Foundation, Norwich, UK

    Google Scholar 

  • Hori T, Horiguchi M, Hayashi A (1984) Biochemistry of natural C-P compounds. Japanese Association for Research on the Biosynthesis of C-P Compounds. Maruzen, Kyoto

    Google Scholar 

  • Imai S, Seto H, Ogawa H, Satoh A, Otake N (1985) Studies on the biosynthesis of fosfomycin. Conversion of 2-hydroxyethylphosphonic acid and 2-aminoethylphosphonic acid to fosfomycin. Agric Biol Chem 49:873–874

    Google Scholar 

  • Kamigiri K, Hidaka T, Imai S, Murakami K, Seto H (1992) Studies on the biosynthesis of bialaphos (SF-1293) 12. C-P bond formation mechanism of bialaphos: discovery of a P-methylation enzyme. J Antibiot 45:781–787

    Google Scholar 

  • Katayama N, Thubotani S, Nozaki Y, Harada S, Ono H (1990) Fosfadecin and fosfocytocin, new nucleotide antibiotics produced by bacteria. J Antibiot 43:238–246

    Google Scholar 

  • Kuzuyama T, Hidaka T, Kamigiri K, Imai S, Seto H (1992) Studies on the biosynthesis of fosfomycin. 4. The biosynthetic origin of the methyl group of fosfomycin. J Antibiot 45:1812–1814

    Google Scholar 

  • Kuzuyama T, Hidaka T, Imai S, Seto H (1993) Studies on the biosynthesis of fosfomycin. 5. Cloning of genes for fosfomycin biosynthesis. J Antibiot 46:1478–1480

    Google Scholar 

  • Matsuda J, Okabe S, Hashimoto T, Yamada Y (1991) Molecular cloning of hyoscyamine 6β-hydroxylase, a 2-oxoglutarate-dependent dioxygenase, from cultured roots of Hyoscyamus niger. J Biol Chem 266:9460–9464

    Google Scholar 

  • Nakashita H, Shimazu A, Hidaka T, Seto H (1992) Purification and characterization of phosphoenolpyruvate phosphomutase from Pseudomonas gladioli B-1. J Bacteriol 174:6857–6861

    Google Scholar 

  • Pearson WR, Lipman DJ (1988) Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 85:2444–2448

    Google Scholar 

  • Peretz M, Burstein Y (1989) Amino acid sequence of alcohol dehydrogenase from the thermophilic bacterium Thermoanaerobium brockii. Biochemistry 28:6549–6555

    Google Scholar 

  • Popham DL, Setlow P (1993) Cloning, nucleotide sequence, and regulation of the Bacillus subtilis pbpE operon, which codes for penicillin-binding protein and an apparent amino acid racemase. J Bacteriol 175:2917–2925

    Google Scholar 

  • Preis S, Myers TC, Jense EV (1995) Phosphonic acids. III: Hydroxyl subtituted propylphosphonic acids. J Am Chem Soc 77:6225–6227

    Google Scholar 

  • Seidel HM, Freeman S, Seto H, Knowles JR (1988) Phosphonate biosynthesis: the isolation and characterization of phosphoenolpyruvate phosphomutase, the enzyme responsible for the formation of a carbon-phosphorus bond. Nature 335:457–458

    Google Scholar 

  • Seidel HM, Pompliano DL, Knowles JR (1992) Phosphonate biosynthesis: molecular cloning of the gene for phosphoenolpyruvate mutase from Tetrahymena pyriformis and over expression of the gene product in Escherichia coli. Biochemistry 31:2598–2608

    Google Scholar 

  • Seto H, Hidaka T, Kuzuyama T, Shibahara S, Usui T, Sakanaka O, Imai S (1991) Studies on the biosynthesis of fosfomycin. 2: Conversion of 2-hydroxypropylphosphonic acid to fosfomycin by blocked mutants of Streptomyces wedmorensis. J Antibiot 44:1286–1288

    Google Scholar 

  • Shoji J, Kato T, Hinoo H, Hattori T, Hirooka K, Matsumoto K, Tanimoto T, Kondo E (1986) Production of fosfomycin (phosphonomycin) by Pseudomonas syringae. J Antibiot 39:1011–1013

    Google Scholar 

  • Thompson CJ, Ward JM, Hopwood DA (1982) Cloning of antibiotic resistance and nutritional genes in Streptomyces. J Bacteriol 151:668–677

    Google Scholar 

  • Vieira J, Messing J (1987) Production of single-stranded plasmid DNA. Methods Enzymol 153:3–11

    Google Scholar 

  • Walker CJ, Mansfield KE, Rezzano IN, Hanamoto CM, Smith KM, Castelfranco PA (1988) The magnesium-protoporphyrin IX (oxidative) cyclase system. Studies on the mechanism and specificity of the reaction sequence. Biochem J 255:685–692

    Google Scholar 

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Communicated by A. Kondorosi

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Hidaka, T., Goda, M., Kuzuyama, T. et al. Cloning and nucleotide sequence of fosfomycin biosynthetic genes of Streptomyces wedmorensis . Molec. Gen. Genet. 249, 274–280 (1995). https://doi.org/10.1007/BF00290527

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  • DOI: https://doi.org/10.1007/BF00290527

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