Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
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Cloning and Characterization of a Heterologous Gene Stimulating Antibiotic Production in Streptomyces lividans TK-24

  • Kwon, Hyung-Jin (Department of Biological Science and Institute of Bioscience & Biotechnology, Myong Ji University, Yongin) ;
  • Lee, Seung-Soo (Department of Biological Science and Institute of Bioscience & Biotechnology, Myong Ji University, Yongin) ;
  • Hong, Soon-Kwang (Department of Biological Science and Institute of Bioscience & Biotechnology, Myong Ji University, Yongin) ;
  • Park, Uhn-Mee (Division of Life Science, University of Suwon) ;
  • Suh, Joo-Won (Department of Biological Science and Institute of Bioscience & Biotechnology, Myong Ji University, Yongin)
  • Published : 1999.06.01
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Abstract

Genetic determinant for the secondary metabolism was studied in heterologous expression in Streptomyces lividans TK-24 using Streptomyces griseus ATCC 10137 as a donor strain. Chromosomal DNA of S. griseus was ligated into the high-copy number Streptomyces shuttle plasmid, pWHM3, and introduced into S. lividans TK-24. A plasmid clone with 4.3-kb BamHI DNA of S. griseus (pMJJ201) was isolated by detecting for stimulatory effect on actinorhodin production by visual inspection. The 4.3-kb BamHI DNA was cloned into pWHM3 under the control of the strong constitutive ermEp promoter in both directions (pMJJ202); ermEp promoter-mediated transcription for coding sequence reading right to left: pMJJ203; ermEp promoter-mediated transcription for coding sequence reading left to right) and reintroduced into S. lividans TK-24. The production of actinorhodin was markedly stimulated due to introduction of pMJJ202 on regeneration agar. The introduction of pMJJ202 also stimulated production of actinorhodin and undecylproidigiosin in submerged culture employing the actinorhodin production medium. Introduction of pMJJ203 resulted in a marked decrease of production of the two pigments. Nucleotide sequence analysis of the 4.3-kb region revealed three coding sequences: two coding sequences reading left to right, ORF1 and ORF2, one coding sequence reading right to left, ORF3. Therefore, it was suggested that the ORF3 product was responsible for the stimulation of antibiotic production. The C-terminal region of ORF3 product showed a local alignment with Myb-related transcriptional factors, which implicated that the ORF3 product might be a novel DNA-binding protein related to the regulation of secondary metabolism in Streptomyces.

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References

  1. J. Mol. Biol. v.215 Basic local alignment search tool Altschul, S.F.;W. Gish;W. Miller;E.W. Myers;D.J. Lipman
  2. Trends Biotechnol. v.13 Signal transduction and secondary metabolism: Prospects for controlling productivity Beppu, T.
  3. Gene v.30 The relationship between base composition and codon usage in bacterial genes and its use for the simple and reliable identification of protein-coding sequences Bibb, M.J.;P.R. Findlay;M.W. Johnson
  4. Gene v.38 Cloning and analysis of the prmoter region of the erythromycin-resistance gene(remE)of Streptomyces erythraeus Bibb, M.J.;G.R. Janssen;J.M. Ward
  5. Microbiology v.142 Clowirth Prize Lecture. The regulation of antibiotic production in Streptomyces coelicolor A3(2) Bibb, M.
  6. Nature v.332 70K heat shock related proteins stimulate protein translocation into microsomes Chirico, W.J.;M.G. Water;G. Blobel
  7. Nature v.332 A subfamily of stress proteins facilitates translocation of secretory and mitochondrial precursor polypeptides Deshaies, R.J.;B.D. Koch;M. Werner-Washburne;E.A. Craig;R. Schekman
  8. Cell v.66 The act cluster contains regulatory and antibiotic export genes, direct targets for translational control by the bldA tRNA gene of Streptomyces Fernandez-Moreno, M.A.;J.L. Caballero;D.A. Hopwood;F. Malpartida
  9. J. Bacteriol. v.174 abaA. A new pleiotropic regulatory locus for antibiotic production in Streptomyces coelicolor Fernandez-Moreno, M.A.;A.J. Martin-Triana;E. Martinez;J. Niemi;H.M. Kieser;D.A. Hopwood;F. Malpartida
  10. Mol. Microbiol. v.7 Stationary-phase production of the antibiotics actinorhodin in Streptomyces coelicolor A3(2) is transcriptionally regulated Gramajo, H.C.;E. Takano;M.J. Bibb
  11. J. Ferment. Bioeng. v.77 Cloning and nucleotide sequence of a hsp70 gene form Streptomyces griseus Hatada, Y.;H. Shinkawa;K. Kawamoto;H. Kinashi;O. Nimi
  12. J. Gen. Microbiol. v.136 Pigment antibiotic production by Streptomyces coelicolor A3(2). kinetics and the influence of nutrients Hobbs, G.;C. M. Frazer;D.C.J. Gardner;F. Flett;S.G. Oliver
  13. J. Bacteriol. v.173 Phosphorylation of the AfsR product, a global regulatory protein for secondary-metabolite formation in Streptomyces coelicolor A3(2) Hong, S.-K.;M. Kito;T. Beppu;S. Horinouchi
  14. A laboratory Manual Genetic manipulation in Streptomyces Hopwood D.A.;M. J. Bibb;K.F. Chater;T. Kieser;C.J. Bruton;H.M. Kieser;D.J. Lydiate;C.P. Smith;J.M. Ward;H. Schrempf
  15. Gene cloning volume; A practical approach(2nd ed.) Gene cloning in Streptomyces Hunter, I.S.;Glover D.M.(ed.)
  16. J. Bacteriol. v.174 A putative two-component regulatory system involved in secondary metabolism in Streptomyces spp Ishizuka, H.;S. Horinouchi;H.M. Kieser;D.A. Hopwood;T. Beppu
  17. Can. J. Microbiol. v.41 Physiologocal control of trophophase-idiophase separation in streptomycetes cultures producing secondary metabolites Liao, X.;L.C. Vining;J.L. Doull
  18. Annu. Rev. Microbiol. v.43 Organization and expression of genes involved in the biosynthesis of antibiotics and other secondary metabolites Martin, J.F.;P.Liras
  19. Gene v.146 Phosphorylation of the AfsR protein involved in secondary metabolism in Streptomyces species by a eukaryotic-type protein kinase Matsumoto, A.;S.-K. Hong;H. Ishizuka;S. Horinouchi;T. Beppu
  20. J. Bacteriol. v.172 Nucleotide sequence and transcriptional analysis of the redD locus of Streptomyces coelicolor A3(2) Narva, K.E.;J.S. Feitelson
  21. Proc. Natl. Acad. Sci. USA v.85 Improved tools for biological sequence comparison Pearson, W.R.;D.J. Lipman
  22. FEMS Microbiol. Lett. v.127 Activation of the actinorhodin biosynthetic pathway in Streptomyces lividans Romero, N.M.;R.P. Mellado
  23. Molecular Cloning: A Laboratory manual Sambrook, J.;E.F. Fritch;T. Maniatis
  24. Proc. Natl. Acad. Sci. USA v.74 DNA sequencing with chain-terminating inhibitors Sanger, F.;S. Nicklen;A.R. Coulson
  25. FEMS Microbiol. Lett. v.147 abaB, a putative regulator for secondary metabolism in Streptomyces Scheu, A.-K.;E. Mart nez;J. Soliveri;F. Malpartida
  26. Regulation of secondary metabolism in Actinomycetes Structural organization and regulation of antibiotic biosynthesis and resistance genes in actionmycetes Seno, E.T.;R.H. Baltz;S. Shapiro(ed.)
  27. Microbiol. Rev. v.53 Protein phosphorylation and regulation of adaptive responses in bacteria Stock, J.B.;A.J. Ninfa;A.M. Stock
  28. Mol. Microbiol. v.6 Transcriptional regulation of the redD transcriptional activator gene accounts for growthphase dependent production of the natibiotic undecylprodigiosin in Streptomyces coelicolor A3(2) Takano, E.;H.C. Gramajo;E. Strauch;N. Andres;J. White;M.J. Bibb
  29. Gene v.20 Physical analysis of antibiotic resistance genes from Streptomyces and their use in vector construction Thompson, C.J.;T. Kieser;J.M. Ward;D.A. Hopwood
  30. FEMS Microbiol lett v.144 Expression of the Streptomyces coelicolor A3(2) ptpA gene encoding a phosphotyrosine protein phosphatase leads to overproduction of secondary metabolites in S Umeyama, T.;Y. Tanabe;B.D. Aigle;S. Horinouchi
  31. J. Bacteriol. v.171 Cloning of genes governing the deoxysugar portion of the erythromycin biosynthetic pathway in Saccharopolyspora erythraea(Streptomyces erythreus) Vara, J.A.;M. Lewandowska-Skarbek;Y.G. Wang;S. Donadio;C.R. Hutchinson
  32. Gene v.33 Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and uPC19 vectors Yanisch-Perron, C.;J. Vieira;J. Messing
  33. J. Bacteriol. v.179 bldA dependence of undecylprodigiosin production in Streptomyces coelicolor A3(2) involves a pathway-specific regulatory cascade White, J.;M. Bibb
  34. Methods Enzymol. v.266 Analysis of compositionally biased regions in sequence databases Wootton, J.C.;S. Federhen