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. 1995 Nov;177(21):6126–6136. doi: 10.1128/jb.177.21.6126-6136.1995

Cloning and characterization of a polyketide synthase gene from Streptomyces fradiae Tü2717, which carries the genes for biosynthesis of the angucycline antibiotic urdamycin A and a gene probably involved in its oxygenation.

H Decker 1, S Haag 1
PMCID: PMC177452  PMID: 7592377

Abstract

A DNA fragment was cloned as cosmid purd8, which encodes a polyketide synthase involved in the production of the angucycline antibiotic urdamycin from Streptomyces fradiae Tü2717. Deletion of the polyketide synthase genes from the chromosome abolished urdamycin production. In addition, purd8 conferred urdamycin resistance on introduction into Streptomyces lividans TK24. Sequence analysis of 5.7 kb of purd8 revealed six open reading frames transcribed in the same direction. The deduced amino acid sequences of the six open reading frames strongly resemble proteins from known type II polyketide synthase gene clusters: a ketoacyl synthase, a chain length factor, an acyl carrier protein, a ketoreductase, a cyclase, and an oxygenase. Heterologous expression of the urdamycin genes encoding a ketoacyl synthase and a chain length factor in Streptomyces glaucescens tetracenomycin C-nonproducing mutants impaired in either the TcmK ketoacyl synthase or TcmL chain length factor resulted in the production of tetracenomycin C. Heterologous expression of a putative oxygenase gene from the urdamycin gene cluster in S. glaucescens GLA.O caused production of the hybrid antibiotic 6-hydroxy tetracenomycin C.

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Selected References

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  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. 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. Bartel P. L., Zhu C. B., Lampel J. S., Dosch D. C., Connors N. C., Strohl W. R., Beale J. M., Jr, Floss H. G. Biosynthesis of anthraquinones by interspecies cloning of actinorhodin biosynthesis genes in streptomycetes: clarification of actinorhodin gene functions. J Bacteriol. 1990 Sep;172(9):4816–4826. doi: 10.1128/jb.172.9.4816-4826.1990. [DOI] [PMC free article] [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. Bibb M. J., Cohen S. N. Gene expression in Streptomyces: construction and application of promoter-probe plasmid vectors in Streptomyces lividans. Mol Gen Genet. 1982;187(2):265–277. doi: 10.1007/BF00331128. [DOI] [PubMed] [Google Scholar]
  6. Bibb M. J., Janssen G. R., Ward J. M. Cloning and analysis of the promoter region of the erythromycin resistance gene (ermE) of Streptomyces erythraeus. Gene. 1985;38(1-3):215–226. doi: 10.1016/0378-1119(85)90220-3. [DOI] [PubMed] [Google Scholar]
  7. Bibb M. J., Sherman D. H., Omura S., Hopwood D. A. Cloning, sequencing and deduced functions of a cluster of Streptomyces genes probably encoding biosynthesis of the polyketide antibiotic frenolicin. Gene. 1994 May 3;142(1):31–39. doi: 10.1016/0378-1119(94)90351-4. [DOI] [PubMed] [Google Scholar]
  8. Blanco G., Pereda A., Brian P., Méndez C., Chater K. F., Salas J. A. A hydroxylase-like gene product contributes to synthesis of a polyketide spore pigment in Streptomyces halstedii. J Bacteriol. 1993 Dec;175(24):8043–8048. doi: 10.1128/jb.175.24.8043-8048.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Decker H., Hutchinson C. R. Transcriptional analysis of the Streptomyces glaucescens tetracenomycin C biosynthesis gene cluster. J Bacteriol. 1993 Jun;175(12):3887–3892. doi: 10.1128/jb.175.12.3887-3892.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Decker H., Motamedi H., Hutchinson C. R. Nucleotide sequences and heterologous expression of tcmG and tcmP, biosynthetic genes for tetracenomycin C in Streptomyces glaucescens. J Bacteriol. 1993 Jun;175(12):3876–3886. doi: 10.1128/jb.175.12.3876-3886.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Drautz H., Zähner H., Rohr J., Zeeck A. Metabolic products of microorganisms. 234. Urdamycins, new angucycline antibiotics from Streptomyces fradiae. I. Isolation, characterization and biological properties. J Antibiot (Tokyo) 1986 Dec;39(12):1657–1669. doi: 10.7164/antibiotics.39.1657. [DOI] [PubMed] [Google Scholar]
  14. Eggink G., Engel H., Vriend G., Terpstra P., Witholt B. Rubredoxin reductase of Pseudomonas oleovorans. Structural relationship to other flavoprotein oxidoreductases based on one NAD and two FAD fingerprints. J Mol Biol. 1990 Mar 5;212(1):135–142. doi: 10.1016/0022-2836(90)90310-I. [DOI] [PubMed] [Google Scholar]
  15. Fernández-Moreno M. A., Martínez E., Boto L., Hopwood D. A., Malpartida F. Nucleotide sequence and deduced functions of a set of cotranscribed genes of Streptomyces coelicolor A3(2) including the polyketide synthase for the antibiotic actinorhodin. J Biol Chem. 1992 Sep 25;267(27):19278–19290. [PubMed] [Google Scholar]
  16. Grimm A., Madduri K., Ali A., Hutchinson C. R. Characterization of the Streptomyces peucetius ATCC 29050 genes encoding doxorubicin polyketide synthase. Gene. 1994 Dec 30;151(1-2):1–10. doi: 10.1016/0378-1119(94)90625-4. [DOI] [PubMed] [Google Scholar]
  17. Hallam S. E., Malpartida F., Hopwood D. A. Nucleotide sequence, transcription and deduced function of a gene involved in polyketide antibiotic synthesis in Streptomyces coelicolor. Gene. 1988 Dec 30;74(2):305–320. doi: 10.1016/0378-1119(88)90165-5. [DOI] [PubMed] [Google Scholar]
  18. Han L., Yang K., Ramalingam E., Mosher R. H., Vining L. C. Cloning and characterization of polyketide synthase genes for jadomycin B biosynthesis in Streptomyces venezuelae ISP5230. Microbiology. 1994 Dec;140(Pt 12):3379–3389. doi: 10.1099/13500872-140-12-3379. [DOI] [PubMed] [Google Scholar]
  19. Hong Y. S., Hwang C. K., Hong S. K., Kim Y. H., Lee J. J. Molecular cloning and characterization of the aklavinone 11-hydroxylase gene of Streptomyces peucetius subsp. caesius ATCC 27952. J Bacteriol. 1994 Nov;176(22):7096–7101. doi: 10.1128/jb.176.22.7096-7101.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hopwood D. A., Malpartida F., Kieser H. M., Ikeda H., Duncan J., Fujii I., Rudd B. A., Floss H. G., Omura S. Production of 'hybrid' antibiotics by genetic engineering. Nature. 1985 Apr 18;314(6012):642–644. doi: 10.1038/314642a0. [DOI] [PubMed] [Google Scholar]
  21. Katz L., Donadio S. Polyketide synthesis: prospects for hybrid antibiotics. Annu Rev Microbiol. 1993;47:875–912. doi: 10.1146/annurev.mi.47.100193.004303. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. McDaniel R., Ebert-Khosla S., Fu H., Hopwood D. A., Khosla C. Engineered biosynthesis of novel polyketides: influence of a downstream enzyme on the catalytic specificity of a minimal aromatic polyketide synthase. Proc Natl Acad Sci U S A. 1994 Nov 22;91(24):11542–11546. doi: 10.1073/pnas.91.24.11542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. McDaniel R., Ebert-Khosla S., Hopwood D. A., Khosla C. Engineered biosynthesis of novel polyketides. Science. 1993 Dec 3;262(5139):1546–1550. doi: 10.1126/science.8248802. [DOI] [PubMed] [Google Scholar]
  25. Motamedi H., Wendt-Pienkowski E., Hutchinson C. R. Isolation of tetracenomycin C-nonproducing Streptomyces glaucescens mutants. J Bacteriol. 1986 Aug;167(2):575–580. doi: 10.1128/jb.167.2.575-580.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Niemi J., Mäntsälä P. Nucleotide sequences and expression of genes from Streptomyces purpurascens that cause the production of new anthracyclines in Streptomyces galilaeus. J Bacteriol. 1995 May;177(10):2942–2945. doi: 10.1128/jb.177.10.2942-2945.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Orser C. S., Lange C. C., Xun L., Zahrt T. C., Schneider B. J. Cloning, sequence analysis, and expression of the Flavobacterium pentachlorophenol-4-monooxygenase gene in Escherichia coli. J Bacteriol. 1993 Jan;175(2):411–416. doi: 10.1128/jb.175.2.411-416.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Perkins E. J., Gordon M. P., Caceres O., Lurquin P. F. Organization and sequence analysis of the 2,4-dichlorophenol hydroxylase and dichlorocatechol oxidative operons of plasmid pJP4. J Bacteriol. 1990 May;172(5):2351–2359. doi: 10.1128/jb.172.5.2351-2359.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Richardson M. A., Kuhstoss S., Solenberg P., Schaus N. A., Rao R. N. A new shuttle cosmid vector, pKC505, for streptomycetes: its use in the cloning of three different spiramycin-resistance genes from a Streptomyces ambofaciens library. Gene. 1987;61(3):231–241. doi: 10.1016/0378-1119(87)90187-9. [DOI] [PubMed] [Google Scholar]
  30. Rohr J., Beale J. M., Floss H. G. Urdamycins, new angucycline antibiotics from Streptomyces fradiae. IV. Biosynthetic studies of urdamycins A-D. J Antibiot (Tokyo) 1989 Jul;42(7):1151–1157. doi: 10.7164/antibiotics.42.1151. [DOI] [PubMed] [Google Scholar]
  31. Rohr J., Thiericke R. Angucycline group antibiotics. Nat Prod Rep. 1992 Apr;9(2):103–137. doi: 10.1039/np9920900103. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Shen B., Hutchinson C. R. Tetracenomycin F1 monooxygenase: oxidation of a naphthacenone to a naphthacenequinone in the biosynthesis of tetracenomycin C in Streptomyces glaucescens. Biochemistry. 1993 Jul 6;32(26):6656–6663. doi: 10.1021/bi00077a019. [DOI] [PubMed] [Google Scholar]
  34. Shen B., Hutchinson C. R. Tetracenomycin F2 cyclase: intramolecular aldol condensation in the biosynthesis of tetracenomycin C in Streptomyces glaucescens. Biochemistry. 1993 Oct 19;32(41):11149–11154. doi: 10.1021/bi00092a026. [DOI] [PubMed] [Google Scholar]
  35. 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]
  36. Summers R. G., Wendt-Pienkowski E., Motamedi H., Hutchinson C. R. Nucleotide sequence of the tcmII-tcmIV region of the tetracenomycin C biosynthetic gene cluster of Streptomyces glaucescens and evidence that the tcmN gene encodes a multifunctional cyclase-dehydratase-O-methyl transferase. J Bacteriol. 1992 Mar;174(6):1810–1820. doi: 10.1128/jb.174.6.1810-1820.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Summers R. G., Wendt-Pienkowski E., Motamedi H., Hutchinson C. R. The tcmVI region of the tetracenomycin C biosynthetic gene cluster of Streptomyces glaucescens encodes the tetracenomycin F1 monooxygenase, tetracenomycin F2 cyclase, and, most likely, a second cyclase. J Bacteriol. 1993 Dec;175(23):7571–7580. doi: 10.1128/jb.175.23.7571-7580.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Tabor S., Richardson C. C. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074–1078. doi: 10.1073/pnas.82.4.1074. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Thompson C. J., Kieser T., Ward J. M., Hopwood D. A. Physical analysis of antibiotic-resistance genes from Streptomyces and their use in vector construction. Gene. 1982 Nov;20(1):51–62. doi: 10.1016/0378-1119(82)90086-5. [DOI] [PubMed] [Google Scholar]
  40. Turner D. H., Sugimoto N., Jaeger J. A., Longfellow C. E., Freier S. M., Kierzek R. Improved parameters for prediction of RNA structure. Cold Spring Harb Symp Quant Biol. 1987;52:123–133. doi: 10.1101/sqb.1987.052.01.017. [DOI] [PubMed] [Google Scholar]
  41. Vara J., Lewandowska-Skarbek M., Wang Y. G., Donadio S., Hutchinson C. R. Cloning of genes governing the deoxysugar portion of the erythromycin biosynthesis pathway in Saccharopolyspora erythraea (Streptomyces erythreus). J Bacteriol. 1989 Nov;171(11):5872–5881. doi: 10.1128/jb.171.11.5872-5881.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Weber W., Zähner H., Siebers J., Schröder K., Zeeck A. Stoffwechselprodukte von Mikroorganismen. 175. Mitteilung. Tetracenomycin C. Arch Microbiol. 1979 May;121(2):111–116. doi: 10.1007/BF00689973. [DOI] [PubMed] [Google Scholar]
  43. Wierenga R. K., Terpstra P., Hol W. G. Prediction of the occurrence of the ADP-binding beta alpha beta-fold in proteins, using an amino acid sequence fingerprint. J Mol Biol. 1986 Jan 5;187(1):101–107. doi: 10.1016/0022-2836(86)90409-2. [DOI] [PubMed] [Google Scholar]
  44. 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]

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