1887

Abstract

ATCC 11455 produces the antifungal polyene antibiotic nystatin containing the deoxysugar moiety mycosamine. Part of the deoxythymidyl diphosphate (TDP)-glucose dehydratase gene () known to be involved in deoxysugar biosynthesis was amplified by PCR from genomic DNA of ATCC 11455. A gene library for was made and screened with the probe. Several overlapping phage clones covering about 30 kb of the genome were physically mapped. A partial DNA sequencing analysis of this region resulted in the identification of several putative genes typical of macrolide antibiotic biosynthetic gene clusters. A gene-transfer system for has been established, and gene deletion or disruption experiments within the putative biosynthetic gene cluster were performed. All of the knock-out mutants retained the ability to produce nystatin, suggesting that the identified gene cluster is not involved in biosynthesis of this antibiotic. Culture extracts from the wild-type strain and three knock-out mutants were analysed by TLC followed by a bioassay against . Two antibacterial compounds were found to be synthesized by the wild-type strain while only one was produced by the mutants. This provided evidence for the involvement of the identified gene cluster in the biosynthesis of a presumably novel antibacterial macrolide antibiotic in .

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-146-3-611
2000-03-01
2024-12-04
Loading full text...

Full text loading...

/deliver/fulltext/micro/146/3/1460611a.html?itemId=/content/journal/micro/10.1099/00221287-146-3-611&mimeType=html&fmt=ahah

References

  1. Bierman M., Logan R., O’Brien K., Seno E. T., Rao R. N., Schoner B. E. 1992; Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp. Gene 116:43–49 [CrossRef]
    [Google Scholar]
  2. Brown R., Hazen E. L. 1953; Phalamycin, an antibacterial agent produced by a Streptomyces noursei variant. Antimicrob Agents Chemother 8:818–821
    [Google Scholar]
  3. Decker H., Gaisser S., Pelzer S., Schneider P., Westrich L., Wohlleben W., Bechthold A. 1996; A general approach for cloning and characterizing dNTP-glucose dehydratase genes from actinomycetes. FEMS Microbiol Lett 141:195–201 [CrossRef]
    [Google Scholar]
  4. Devereux J., Haeberli P., Smithies O. 1984; A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395 [CrossRef]
    [Google Scholar]
  5. Donadio S., Staver M. J. 1993; IS1136, an insertion element in the erythromycin gene cluster of Saccharopolyspora erythraea. Gene 126:147–151 [CrossRef]
    [Google Scholar]
  6. Donadio S., Staver M. J., McAlpine J. B., Swanson S. J., Katz L. 1991; Modular organization of genes required for complex polyketide biosynthesis. Science 252:675–679 [CrossRef]
    [Google Scholar]
  7. Flett F., Mersinias V., Smith C. P. 1997; High efficiency conjugal transfer of plasmid DNA from Escherichia coli to methyl DNA-restricting streptomycetes. FEMS Microbiol Lett 155:223–229 [CrossRef]
    [Google Scholar]
  8. Gaisser S., Bohm G. A., Cortes J., Leadlay P. F. 1997; Analysis of seven genes from the eryAIeryK region of the erythromycin biosynthetic gene cluster in Saccharopolyspora erythraea. Mol Gen Genet 256:239–251 [CrossRef]
    [Google Scholar]
  9. Hopwood D. A. 1997; Genetic contributions to understanding polyketide synthases. Chem Rev 97:2465–2497 [CrossRef]
    [Google Scholar]
  10. Hopwood D. A., Bibb M. J., Chater K. F.7 other authors 1985 Genetic Manipulation of Streptomyces: a Laboratory Manual Norwich: John Innes Foundation;
    [Google Scholar]
  11. Inouye M., Suzuki H., Takada Y., Muto N., Horinouchi S., Beppu T. 1994; A gene encoding mycinamicin III O-methyltransferase from Micromonospora griseorubida. Gene 141:121–124 [CrossRef]
    [Google Scholar]
  12. Kaluza K., Hahn M., Hennecke H. 1985; Repeated sequences similar to insertion elements clustered around the nif region of the Rhizobium japonicum genome. J Bacteriol 162:535–542
    [Google Scholar]
  13. Katz L., Donadio S. 1995; Macrolides. In Genetics and Biochemistry of Antibiotic Production pp. 385–420Edited by Vining L. C., Stuttard C. Boston, MA: Butterworth-Heinemann;
    [Google Scholar]
  14. Koplin R., Wang G., Hotte B., Priefer U. B., Puhler A. 1993; A 3·9 kb DNA region of Xanthomonas campestris pv. campestris that is necessary for lipopolysaccharide production encodes a set of enzymes involved in the synthesis of dTDP-rhamnose. J Bacteriol 175:7786–7792
    [Google Scholar]
  15. Kuhstoss S., Huber M., Turner J. R., Paschal J. W., Rao R. N. 1996; Production of a novel polyketide through the construction of a hybrid polyketide synthase. Gene 183:231–236 [CrossRef]
    [Google Scholar]
  16. Liu H. W., Thorson J. S. 1994; Pathways and mechanisms in the biogenesis of novel deoxysugars by bacteria. Annu Rev Microbiol 48:223–256 [CrossRef]
    [Google Scholar]
  17. Lombo F., Siems K., Brana A. F., Mendez C., Bindseil K., Salas J. A. 1997; Cloning and insertional inactivation of Streptomyces argillaceus genes involved in the earliest steps of biosynthesis of the sugar moieties of the antitumour polyketide mithramycin. J Bacteriol 179:3354–3357
    [Google Scholar]
  18. MacNeil D. J., Gewain K. M., Ruby C. L., Dezeny G., Gibbons P. H., MacNeil T. 1992; Analysis of Streptomyces avermitilis genes required for avermectin biosynthesis utilizing a novel integration vector. Gene 111:61–68 [CrossRef]
    [Google Scholar]
  19. Matsushima P., Baltz R. H. 1994; Transformation of Saccharopolyspora spinosa protoplasts with plasmid DNA modified in vitro to avoid host restriction. Microbiology 140:139–143 [CrossRef]
    [Google Scholar]
  20. Mazodier P., Petter R., Thompson C. 1989; Intergeneric conjugation between Escherichia coli and Streptomyces species. J Bacteriol 171:3583–3585
    [Google Scholar]
  21. Olano C., Rodriguez A. M., Mendez C., Salas J. A. 1995; A second ABC transporter is involved in oleandomycin resistance and its secretion by Streptomyces antibioticus. Mol Microbiol 16:333–343 [CrossRef]
    [Google Scholar]
  22. Roszkowski J., Kotiuszko D., Rafalski A., Morawska H., Raczynska-Bojanowska K. 1972; Characteristics of mutants of Streptomyces noursei var. polifungini producing antibiotics. Acta Microbiol Pol B 4:9–22
    [Google Scholar]
  23. Ruan X., Pereda A., Stassi D. L.8 other authors 1997; Acyltransferase domain substitutions in erythromycin polyketide synthase yield novel erythromycin derivatives. J Bacteriol 179:6416–6425
    [Google Scholar]
  24. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  25. Schwecke T., Aparicio J. F., Molnar I.10 other authors 1995; The biosynthetic gene cluster for the polyketide immunosuppressant rapamycin. Proc Natl Acad Sci USA 92:7839–7843 [CrossRef]
    [Google Scholar]
  26. Sezonov G., Blanc V., Bamas-Jacques N., Friedmann A., Pernodet J.-L., Guerineau M. 1997; Complete conversion of antibiotic precursor to pristinamycin IIA by overexpresion of Streptomyces pristinaespiralis biosynthetic genes. Nat Biotechnol 15:349–355 [CrossRef]
    [Google Scholar]
  27. Solenberg P. J., Matsushima P., Stack D. R., Wilkie S. C., Thompson R. C., Baltz R. H. 1997; Production of hybrid glycopeptide antibiotics in vitro and in Streptomyces toyocaensis. Chem Biol 4:195–202 [CrossRef]
    [Google Scholar]
  28. Tsukioka Y., Yamashita Y., Oho T., Nakano Y., Koga T. 1997; Biological function of the dTDP-rhamnose synthesis pathway in Streptococcus mutans. J Bacteriol 179:1126–1134
    [Google Scholar]
  29. Vara J., Lewandowska-Skarbek M., Wang Y.-G., Donadio S., Hutchinson C. R. 1989; Cloning of genes governing the deoxysugar portion of the erythromycin biosynthesis pathway in Saccharopolyspora erythraea (Streptomyces erythraeus). J Bacteriol 171:5872–5881
    [Google Scholar]
  30. Vogtli M., Hutter R. 1987; Characterisation of the hydroxystreptomycin phosphotransferase gene (sph) of Streptomyces glaucescens: nucleotide sequence and promoter analysis. Mol Gen Genet 208:195–203 [CrossRef]
    [Google Scholar]
  31. Weber J. M., Leung J. O., Swanson S. J., Idler K. B., McAlpine J. B. 1991; An erythromycin derivative produced by targeted gene disruption in Saccharopolyspora erythraea. Science 252:114–117 [CrossRef]
    [Google Scholar]
  32. Wohlleben W., Muth G. 1993; Streptomyces plasmid vectors. In Plasmids, a Practical Approach pp. 147–175Edited by Hardy K. G. Oxford, New York & Tokyo: IRL Press;
    [Google Scholar]
  33. Ylihonko K., Tuikkanen J., Jussila S., Cong L., Mantsala P. 1996; A gene cluster involved in nogalamycin biosynthesis from Streptomyces nogalater: sequence analysis and complementation of early-block mutations in the anthracycline pathway. Mol Gen Genet 251:113–120
    [Google Scholar]
/content/journal/micro/10.1099/00221287-146-3-611
Loading
/content/journal/micro/10.1099/00221287-146-3-611
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error