Skip to main content

Some NLM-NCBI services and products are experiencing heavy traffic, which may affect performance and availability. We apologize for the inconvenience and appreciate your patience. For assistance, please contact our Help Desk at info@ncbi.nlm.nih.gov.

Journal of Bacteriology logoLink to Journal of Bacteriology
. 1997 Dec;179(23):7515–7522. doi: 10.1128/jb.179.23.7515-7522.1997

Identification and characterization of the niddamycin polyketide synthase genes from Streptomyces caelestis.

S J Kakavas 1, L Katz 1, D Stassi 1
PMCID: PMC179704  PMID: 9393718

Abstract

The genes encoding the polyketide synthase (PKS) portion of the niddamycin biosynthetic pathway were isolated from a library of Streptomyces caelestis NRRL-2821 chromosomal DNA. Analysis of 40 kb of DNA revealed the presence of five large open reading frames (ORFs) encoding the seven modular sets of enzymatic activities required for the synthesis of a 16-membered lactone ring. The enzymatic motifs identified within each module were consistent with those predicted from the structure of niddamycin. Disruption of the second ORF of the PKS coding region eliminated niddamycin production, demonstrating that the cloned genes are involved in the biosynthesis of this compound.

Full Text

The Full Text of this article is available as a PDF (637.6 KB).

Selected References

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

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Aparicio J. F., Molnár I., Schwecke T., König A., Haydock S. F., Khaw L. E., Staunton J., Leadlay P. F. Organization of the biosynthetic gene cluster for rapamycin in Streptomyces hygroscopicus: analysis of the enzymatic domains in the modular polyketide synthase. Gene. 1996 Feb 22;169(1):9–16. doi: 10.1016/0378-1119(95)00800-4. [DOI] [PubMed] [Google Scholar]
  4. Bevitt D. J., Cortes J., Haydock S. F., Leadlay P. F. 6-Deoxyerythronolide-B synthase 2 from Saccharopolyspora erythraea. Cloning of the structural gene, sequence analysis and inferred domain structure of the multifunctional enzyme. Eur J Biochem. 1992 Feb 15;204(1):39–49. doi: 10.1111/j.1432-1033.1992.tb16603.x. [DOI] [PubMed] [Google Scholar]
  5. Bierman M., Logan R., O'Brien K., Seno E. T., Rao R. N., Schoner B. E. Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp. Gene. 1992 Jul 1;116(1):43–49. doi: 10.1016/0378-1119(92)90627-2. [DOI] [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. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [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., McAlpine J. B., Sheldon P. J., Jackson M., Katz L. An erythromycin analog produced by reprogramming of polyketide synthesis. Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):7119–7123. doi: 10.1073/pnas.90.15.7119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Gandecha A. R., Large S. L., Cundliffe E. Analysis of four tylosin biosynthetic genes from the tylLM region of the Streptomyces fradiae genome. Gene. 1997 Jan 15;184(2):197–203. doi: 10.1016/s0378-1119(96)00595-1. [DOI] [PubMed] [Google Scholar]
  12. HUBER G., WALLHAEUSSER K. H., FRIES L., STEIGLER A., WEIDENMUELLER H. L. [Niddamycin, a new macrolide antibiotic]. Arzneimittelforschung. 1962 Dec;12:1191–1195. [PubMed] [Google Scholar]
  13. Haydock S. F., Aparicio J. F., Molnár I., Schwecke T., Khaw L. E., König A., Marsden A. F., Galloway I. S., Staunton J., Leadlay P. F. Divergent sequence motifs correlated with the substrate specificity of (methyl)malonyl-CoA:acyl carrier protein transacylase domains in modular polyketide synthases. FEBS Lett. 1995 Oct 30;374(2):246–248. doi: 10.1016/0014-5793(95)01119-y. [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. Kuhstoss S., Huber M., Turner J. R., Paschal J. W., Rao R. N. Production of a novel polyketide through the construction of a hybrid polyketide synthase. Gene. 1996 Dec 12;183(1-2):231–236. doi: 10.1016/s0378-1119(96)00565-3. [DOI] [PubMed] [Google Scholar]
  16. Lomovskaya N., Fonstein L., Ruan X., Stassi D., Katz L., Hutchinson C. R. Gene disruption and replacement in the rapamycin-producing Streptomyces hygroscopicus strain ATCC 29253. Microbiology. 1997 Mar;143(Pt 3):875–883. doi: 10.1099/00221287-143-3-875. [DOI] [PubMed] [Google Scholar]
  17. Merson-Davies L. A., Cundliffe E. Analysis of five tylosin biosynthetic genes from the tyllBA region of the Streptomyces fradiae genome. Mol Microbiol. 1994 Jul;13(2):349–355. doi: 10.1111/j.1365-2958.1994.tb00428.x. [DOI] [PubMed] [Google Scholar]
  18. O'Hagan D. Biosynthesis of fatty acid and polyketide metabolites. Nat Prod Rep. 1993 Dec;10(6):593–624. doi: 10.1039/np9931000593. [DOI] [PubMed] [Google Scholar]
  19. Oliynyk M., Brown M. J., Cortés J., Staunton J., Leadlay P. F. A hybrid modular polyketide synthase obtained by domain swapping. Chem Biol. 1996 Oct;3(10):833–839. doi: 10.1016/s1074-5521(96)90069-1. [DOI] [PubMed] [Google Scholar]
  20. Omura S., Nakagawa A., Takeshima H., Atusmi K., Miyazawa J., Piriou F., Lukacs G. Letter: Biosynthetic studies using 13C enriched precursors on the 16-membered macrolide antibiotic leucomycin A3. J Am Chem Soc. 1975 Oct 29;97(22):6600–6602. doi: 10.1021/ja00855a065. [DOI] [PubMed] [Google Scholar]
  21. Omura S., Tsuzuki K., Nakagawa A., Lukacs G. Biosynthetic origin of carbons 3 and 4 of leucomycin aglycone. J Antibiot (Tokyo) 1983 May;36(5):611–613. doi: 10.7164/antibiotics.36.611. [DOI] [PubMed] [Google Scholar]
  22. Ruan X., Pereda A., Stassi D. L., Zeidner D., Summers R. G., Jackson M., Shivakumar A., Kakavas S., Staver M. J., Donadio S. Acyltransferase domain substitutions in erythromycin polyketide synthase yield novel erythromycin derivatives. J Bacteriol. 1997 Oct;179(20):6416–6425. doi: 10.1128/jb.179.20.6416-6425.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schwecke T., Aparicio J. F., Molnár I., König A., Khaw L. E., Haydock S. F., Oliynyk M., Caffrey P., Cortés J., Lester J. B. The biosynthetic gene cluster for the polyketide immunosuppressant rapamycin. Proc Natl Acad Sci U S A. 1995 Aug 15;92(17):7839–7843. doi: 10.1073/pnas.92.17.7839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Scrutton N. S., Berry A., Perham R. N. Redesign of the coenzyme specificity of a dehydrogenase by protein engineering. Nature. 1990 Jan 4;343(6253):38–43. doi: 10.1038/343038a0. [DOI] [PubMed] [Google Scholar]
  25. Strohl W. R. Compilation and analysis of DNA sequences associated with apparent streptomycete promoters. Nucleic Acids Res. 1992 Mar 11;20(5):961–974. doi: 10.1093/nar/20.5.961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tuan J. S., Weber J. M., Staver M. J., Leung J. O., Donadio S., Katz L. Cloning of genes involved in erythromycin biosynthesis from Saccharopolyspora erythraea using a novel actinomycete-Escherichia coli cosmid. Gene. 1990 May 31;90(1):21–29. doi: 10.1016/0378-1119(90)90435-t. [DOI] [PubMed] [Google Scholar]
  27. Witkowski A., Rangan V. S., Randhawa Z. I., Amy C. M., Smith S. Structural organization of the multifunctional animal fatty-acid synthase. Eur J Biochem. 1991 Jun 15;198(3):571–579. doi: 10.1111/j.1432-1033.1991.tb16052.x. [DOI] [PubMed] [Google Scholar]
  28. Wright F., Bibb M. J. Codon usage in the G+C-rich Streptomyces genome. Gene. 1992 Apr 1;113(1):55–65. doi: 10.1016/0378-1119(92)90669-g. [DOI] [PubMed] [Google Scholar]
  29. Yamamoto H., Maurer K. H., Hutchinson C. R. Transformation of Streptomyces erythraeus. J Antibiot (Tokyo) 1986 Sep;39(9):1304–1313. doi: 10.7164/antibiotics.39.1304. [DOI] [PubMed] [Google Scholar]
  30. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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

RESOURCES