Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1995 Jul;177(13):3673–3679. doi: 10.1128/jb.177.13.3673-3679.1995

A Sorangium cellulosum (myxobacterium) gene cluster for the biosynthesis of the macrolide antibiotic soraphen A: cloning, characterization, and homology to polyketide synthase genes from actinomycetes.

T Schupp 1, C Toupet 1, B Cluzel 1, S Neff 1, S Hill 1, J J Beck 1, J M Ligon 1
PMCID: PMC177082  PMID: 7601830

Abstract

A 40-kb region of DNA from Sorangium cellulosum So ce26, which contains polyketide synthase (PKS) genes for synthesis of the antifungal macrolide antibiotic soraphen A, was cloned. These genes were detected by homology to Streptomyces violaceoruber genes encoding components of granaticin PKS, thus extending this powerful technique for the identification of bacterial PKS genes, which has so far been applied only to actinomycetes, to the gram-negative myxobacteria. Functional analysis by gene disruption has indicated that about 32 kb of contiguous DNA of the cloned region contains genes involved in soraphen A biosynthesis. The nucleotide sequence of a 6.4-kb DNA fragment, derived from the region with homology to granaticin PKS genes, was determined. Analysis of this sequence has revealed the presence of a single large open reading frame beginning and ending outside the 6.4-kb fragment. The deduced amino acid sequence indicates the presence of a domain with a high level of similarity to beta-ketoacyl synthases that are involved in polyketide synthesis. Other domains with high levels of similarity to regions of known polyketide biosynthetic functions were identified, including those for acyl transferase, acyl carrier protein, ketoreductase, and dehydratase. We present data which indicate that soraphen A biosynthesis is catalyzed by large, multifunctional enzymes analogous to other bacterial PKSs of type I.

Full Text

The Full Text of this article is available as a PDF (402.3 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amy C. M., Witkowski A., Naggert J., Williams B., Randhawa Z., Smith S. Molecular cloning and sequencing of cDNAs encoding the entire rat fatty acid synthase. Proc Natl Acad Sci U S A. 1989 May;86(9):3114–3118. doi: 10.1073/pnas.86.9.3114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arrowsmith T. J., Malpartida F., Sherman D. H., Birch A., Hopwood D. A., Robinson J. A. Characterisation of actI-homologous DNA encoding polyketide synthase genes from the monensin producer Streptomyces cinnamonensis. Mol Gen Genet. 1992 Aug;234(2):254–264. doi: 10.1007/BF00283846. [DOI] [PubMed] [Google Scholar]
  3. Beck J., Ripka S., Siegner A., Schiltz E., Schweizer E. The multifunctional 6-methylsalicylic acid synthase gene of Penicillium patulum. Its gene structure relative to that of other polyketide synthases. Eur J Biochem. 1990 Sep 11;192(2):487–498. doi: 10.1111/j.1432-1033.1990.tb19252.x. [DOI] [PubMed] [Google Scholar]
  4. Bibb M. J., Biró S., Motamedi H., Collins J. F., Hutchinson C. R. Analysis of the nucleotide sequence of the Streptomyces glaucescens tcmI genes provides key information about the enzymology of polyketide antibiotic biosynthesis. EMBO J. 1989 Sep;8(9):2727–2736. doi: 10.1002/j.1460-2075.1989.tb08414.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  6. Cortes J., Haydock S. F., Roberts G. A., Bevitt D. J., Leadlay P. F. An unusually large multifunctional polypeptide in the erythromycin-producing polyketide synthase of Saccharopolyspora erythraea. Nature. 1990 Nov 8;348(6297):176–178. doi: 10.1038/348176a0. [DOI] [PubMed] [Google Scholar]
  7. Davis N. K., Chater K. F. Spore colour in Streptomyces coelicolor A3(2) involves the developmentally regulated synthesis of a compound biosynthetically related to polyketide antibiotics. Mol Microbiol. 1990 Oct;4(10):1679–1691. doi: 10.1111/j.1365-2958.1990.tb00545.x. [DOI] [PubMed] [Google Scholar]
  8. Donadio S., Katz L. Organization of the enzymatic domains in the multifunctional polyketide synthase involved in erythromycin formation in Saccharopolyspora erythraea. Gene. 1992 Feb 1;111(1):51–60. doi: 10.1016/0378-1119(92)90602-l. [DOI] [PubMed] [Google Scholar]
  9. Donadio S., Staver M. J., McAlpine J. B., Swanson S. J., Katz L. Modular organization of genes required for complex polyketide biosynthesis. Science. 1991 May 3;252(5006):675–679. doi: 10.1126/science.2024119. [DOI] [PubMed] [Google Scholar]
  10. Gerth K., Bedorf N., Irschik H., Höfle G., Reichenbach H. The soraphens: a family of novel antifungal compounds from Sorangium cellulosum (Myxobacteria). I. Soraphen A1 alpha: fermentation, isolation, biological properties. J Antibiot (Tokyo) 1994 Jan;47(1):23–31. doi: 10.7164/antibiotics.47.23. [DOI] [PubMed] [Google Scholar]
  11. Hedges R. W., Matthew M. Acquisition by Escherichia coli of plasmid-borne beta-lactamases normally confined to Pseudomonas spp. Plasmid. 1979 Apr;2(2):269–278. doi: 10.1016/0147-619x(79)90045-3. [DOI] [PubMed] [Google Scholar]
  12. Hohn B., Collins J. A small cosmid for efficient cloning of large DNA fragments. Gene. 1980 Nov;11(3-4):291–298. doi: 10.1016/0378-1119(80)90069-4. [DOI] [PubMed] [Google Scholar]
  13. Hopwood D. A., Khosla C. Genes for polyketide secondary metabolic pathways in microorganisms and plants. Ciba Found Symp. 1992;171:88–112. doi: 10.1002/9780470514344.ch6. [DOI] [PubMed] [Google Scholar]
  14. Hopwood D. A., Sherman D. H. Molecular genetics of polyketides and its comparison to fatty acid biosynthesis. Annu Rev Genet. 1990;24:37–66. doi: 10.1146/annurev.ge.24.120190.000345. [DOI] [PubMed] [Google Scholar]
  15. Jaoua S., Neff S., Schupp T. Transfer of mobilizable plasmids to Sorangium cellulosum and evidence for their integration into the chromosome. Plasmid. 1992 Sep;28(2):157–165. doi: 10.1016/0147-619x(92)90046-d. [DOI] [PubMed] [Google Scholar]
  16. Kakinuma S., Takada Y., Ikeda H., Tanaka H., Omura S., Hopwood D. A. Cloning of large DNA fragments, which hybridize with actinorhodin biosynthesis genes, from kalafungin and nanaomycin A methyl ester producers and identification of genes for kalafungin biosynthesis of the kalafungin producer. J Antibiot (Tokyo) 1991 Sep;44(9):995–1005. doi: 10.7164/antibiotics.44.995. [DOI] [PubMed] [Google Scholar]
  17. Kauppinen S., Siggaard-Andersen M., von Wettstein-Knowles P. beta-Ketoacyl-ACP synthase I of Escherichia coli: nucleotide sequence of the fabB gene and identification of the cerulenin binding residue. Carlsberg Res Commun. 1988;53(6):357–370. doi: 10.1007/BF02983311. [DOI] [PubMed] [Google Scholar]
  18. Malpartida F., Hallam S. E., Kieser H. M., Motamedi H., Hutchinson C. R., Butler M. J., Sugden D. A., Warren M., McKillop C., Bailey C. R. Homology between Streptomyces genes coding for synthesis of different polyketides used to clone antibiotic biosynthetic genes. 1987 Feb 26-Mar 4Nature. 325(6107):818–821. doi: 10.1038/325818a0. [DOI] [PubMed] [Google Scholar]
  19. Murray N. E., Brammar W. J., Murray K. Lambdoid phages that simplify the recovery of in vitro recombinants. Mol Gen Genet. 1977 Jan 7;150(1):53–61. doi: 10.1007/BF02425325. [DOI] [PubMed] [Google Scholar]
  20. Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
  21. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sherman D. H., Malpartida F., Bibb M. J., Kieser H. M., Bibb M. J., Hopwood D. A. Structure and deduced function of the granaticin-producing polyketide synthase gene cluster of Streptomyces violaceoruber Tü22. EMBO J. 1989 Sep;8(9):2717–2725. doi: 10.1002/j.1460-2075.1989.tb08413.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Swan D. G., Rodríguez A. M., Vilches C., Méndez C., Salas J. A. Characterisation of a Streptomyces antibioticus gene encoding a type I polyketide synthase which has an unusual coding sequence. Mol Gen Genet. 1994 Feb;242(3):358–362. doi: 10.1007/BF00280426. [DOI] [PubMed] [Google Scholar]
  24. Vahlensieck H. F., Pridzun L., Reichenbach H., Hinnen A. Identification of the yeast ACC1 gene product (acetyl-CoA carboxylase) as the target of the polyketide fungicide soraphen A. Curr Genet. 1994 Feb;25(2):95–100. doi: 10.1007/BF00309532. [DOI] [PubMed] [Google Scholar]
  25. Varon M., Fuchs N., Monosov M., Tolchinsky S., Rosenberg E. Mutation and mapping of genes involved in production of the antibiotic TA in Myxococcus xanthus. Antimicrob Agents Chemother. 1992 Oct;36(10):2316–2321. doi: 10.1128/aac.36.10.2316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Yu T. W., Bibb M. J., Revill W. P., Hopwood D. A. Cloning, sequencing, and analysis of the griseusin polyketide synthase gene cluster from Streptomyces griseus. J Bacteriol. 1994 May;176(9):2627–2634. doi: 10.1128/jb.176.9.2627-2634.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES