Applied Microbiology and Biotechnology

, Volume 72, Issue 1, pp 145–154 | Cite as

Characterization of Streptomyces MITKK-103, a newly isolated actinomycin X2-producer

  • K. Kurosawa
  • V. P. Bui
  • J. L. VanEssendelft
  • L. B. Willis
  • P. A. Lessard
  • I. Ghiviriga
  • T. G. Sambandan
  • C. K. Rha
  • A. J. Sinskey
Applied Microbial and Cell Physiology
First Online:
Received:
Revised:
Accepted:

Abstract

A new actinomycete strain designated MITKK-103 was isolated from the soil of a flowerpot using a humic acid agar medium. The newly isolated strain was able to produce a large amount of actinomycin X2 even under nonoptimized growing conditions and serves as a promising source of this antibiotic. Actinomycin X2 has higher cytotoxicity toward cultured human leukemia (HL-60) cells than does actinomycin D, and it induces cell death via apoptosis. A nearly complete 16S ribosomal DNA (rDNA) sequence from the isolate was determined and found to have high identity (98.5–100%) with Streptomyces galbus, Streptomyces griseofuscus, and Streptomyces padanus, indicating that MITKK-103 belongs to the genus Streptomyces. The isolate clustered with species belonging to the S. padanus clade in a 16S-rDNA-based phylogenetic tree and showed 75% overall homology to S. padanus ATCC 25646 in DNA–DNA relatedness analysis. Although the growth of the isolate was somewhat different from the three species mentioned, the strain MITKK-103 most closely resembles S. padanus on the basis of the morphological and phenotypic characteristics, phylogenetic analysis, and genotypic data. As such, this is the first report of a strain of S. padanus capable of producing actinomycins.

Keywords

Streptomyces Actinomycin International Streptomyces Project Fungichromin Agar Diffusion Bioassay 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
This is a preview of subscription content, log in to check access.

Notes

Acknowledgements

This work was funded in part by the Government of Malaysia and the Malaysia-MIT Biotechnology Partnership Programme. We would like to thank Dr. Jacqueline Piret for helpful suggestions on the manuscript.

References

  1. Alekhova TA, Novozhilova T (2001) Biosynthesis of polyketide antibiotics by various actinomycin producing Streptomyces species. Prikl Biokhim Mikrobiol 37:309–316PubMedGoogle Scholar
  2. Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402CrossRefPubMedGoogle Scholar
  3. Anderson AS, Wellington EM (2001) The taxonomy of Streptomyces and related genera. Int J Syst Evol Microbiol 51:797–814PubMedGoogle Scholar
  4. Baldacci E, Farina G, Piacenza E, Fabbri G (1968) Description of Streptomyces padanus sp. nov. and emendation of Streptomyces xantophaeus. G Microbiol 16:9–16Google Scholar
  5. Bolognese A, Correale G, Manfra M, Lavecchia A, Mazzoni O, Novellino E, Barone V, Pani A, Tramontano E, La Colla P, Murgioni C, Serra I, Setzu G, Loddo R (2002) Antitumor agents. 1. Synthesis, biological evaluation, and molecular modeling of 5H-pyrido[3,2-a]phenoxazin-5-one, a compound with potent antiproliferative activity. J Med Chem 45:5205–5216CrossRefPubMedGoogle Scholar
  6. Bossi R, Hutter R, Keller-Schierlein W, Neipp L, Zahner H (1958) Metabolic products from actinomycetes. XIV. Actinomycin Z. Helv Chim Acta 41:1645–1652CrossRefGoogle Scholar
  7. Brosius J, Palmer ML, Kennedy PJ, Noller HF (1978) Complete nucleotide sequence of a 16S ribosomal RNA gene from Escherichia coli. Proc Natl Acad Sci U S A 75:4801–4805PubMedCrossRefGoogle Scholar
  8. Castillo UF, Strobel GA, Ford EJ, Hess WM, Porter H, Jensen JB, Albert H, Robison R, Condron MA, Teplow DB, Stevens D, Yaver D (2002) Munumbicins, wide-spectrum antibiotics produced by Streptomyces NRRL 30562, endophytic on Kennedia nigriscans. Microbiology 148:2675–2685PubMedGoogle Scholar
  9. Chen FM, Sha F, Chin KH, Chou SH (2004) The nature of actinomycin D binding to d(AACCAXYG) sequence motifs. Nucleic Acids Res 32:271–277CrossRefPubMedGoogle Scholar
  10. El-Gammal AA (1987) Characterization of an orange-brown pigment-antibiotic produced by Streptomyces viridiviolaceus. Egypt J Microbiol 21:37–42Google Scholar
  11. El-Naggar MYM (1998) Synergistic effect of actinomycin X2 produced by Streptomyces nasri YG62 with other antibiotics. Biomed Lett 58:169–173Google Scholar
  12. Ezaki T, Hashimoto Y, Yabuuchi E (1989) Fluorometric deoxyribonucleic acid–deoxyribonucleic acid hybridization in microdilution wells as an alternative to membrane filter hybridization in which radioisotopes are used to determine genetic relatedness among bacterial strains. Int J Syst Bacteriol 39:224–229Google Scholar
  13. Farber S, D'Angio G, Evans A, Mitus A (2002) Clinical studies of actinomycin D with special reference to Wilms' tumor in children. 1960. J Urol 168:2560–2562; discussion 2563CrossRefPubMedGoogle Scholar
  14. Frommer W (1959) Zur Systematik der actinomycin bildenden streptomyceten. Arch Mikrobiol 32:187–206CrossRefPubMedGoogle Scholar
  15. Green DM (1997) Paediatric oncology update/Wilms' tumour. Eur J Cancer 33:409–418CrossRefPubMedGoogle Scholar
  16. Guo J, Wu T, Bess J, Henderson LE, Levin JG (1998) Actinomycin D inhibits human immunodeficiency virus type 1 minus-strand transfer in in vitro and endogenous reverse transcriptase assays. J Virol 72:6716–6724PubMedGoogle Scholar
  17. Gupta KC, Sobti RR, Chopra IC (1963) Actinomycin produced by a new species of Streptomyces. Hindustan Antibiot Bull 6:12–16Google Scholar
  18. Ha SC, Hong SD (1994) Identification of the actinomycete strain No.1372, a producer of actinomycin X2. Sanop Misaengmul Hakhoechi 22:164–168Google Scholar
  19. Hirata Y, Nakanishi K (1949) Actinomycin J2, a by-product from a strain of Actinomyces. Bull Chem Soc Jpn 22:121–127CrossRefGoogle Scholar
  20. Jacobson E, Dstwald W (1942) The color harmony manual and how to use it. Color Laboratories Division, ChicagoGoogle Scholar
  21. Kataoka M, Ueda K, Kudo T, Seki T, Yoshida T (1997) Application of the variable region in 16S rDNA to create an index for rapid species identification in the genus Streptomyces. FEMS Microbiol Lett 151:249–255CrossRefPubMedGoogle Scholar
  22. Kleeff J, Kornmann M, Sawhney H, Korc M (2000) Actinomycin D induces apoptosis and inhibits growth of pancreatic cancer cells. Int J Cancer 86:399–407CrossRefPubMedGoogle Scholar
  23. Kurzatkowski W (1975) Role of Streptomyces melanochromogenes cell wall and membrane in actinomycin D biosynthesis. Med Dosw Mikrobiol 27:325–336PubMedGoogle Scholar
  24. Lam KS, Gustavson DR, Pirnik DL, Pack E, Bulanhagui C, Mamber SW, Forenza S, Stodieck LS, Klaus DM (2002) The effect of space flight on the production of actinomycin D by Streptomyces plicatus. J Ind Microbiol Biotechnol 29:299–302CrossRefPubMedGoogle Scholar
  25. Malpartida F, Hallam SE, Kieser HM, Motamedi H, Hutchinson CR, Butler MJ, Sugden DA, Warren M, McKillop C, Bailey CR et al (1987) Homology between Streptomyces genes coding for synthesis of different polyketides used to clone antibiotic biosynthetic genes. Nature 325:818–821CrossRefPubMedGoogle Scholar
  26. Marmur J (1961) A procedure for the isolation of deoxyribonucleic acid from micro-organisms. J Mol Biol 3:208–218CrossRefGoogle Scholar
  27. Marroquin AS (1959) Actinomycin A from Streptomyces globosus. Rev Soc Quim Mex 3:19–22Google Scholar
  28. Mauger AB (1980) The actinomycins. Ellis Horwood, EnglandGoogle Scholar
  29. Men'shikov GP, Denisova SI (1962) Isolation of actinomycins from the group Actinomyces fluorescens. Antibiotiki 7:31–32PubMedGoogle Scholar
  30. Mossman T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63CrossRefPubMedGoogle Scholar
  31. Onaka H, Taniguchi S, Igarashi Y, Furumai T (2002) Cloning of the staurosporine biosynthetic gene cluster from Streptomyces sp. TP-A0274 and its heterologous expression in Streptomyces lividans. J Antibiot (Tokyo) 55:1063–1071Google Scholar
  32. Pernodet JL, Boccard F, Alegre MT, Gagnat J, Guerineau M (1989) Organization and nucleotide sequence analysis of a ribosomal RNA gene cluster from Streptomyces ambofaciens. Gene 79:33–46CrossRefPubMedGoogle Scholar
  33. Pirt SJ, Pickett SMA, Pickett AM, Vandamme EJ, Bird CW (1981) Antibiotics from the newly isolated Streptomyces elizabethii. J Chem Technol Biotech 167–177:167–177Google Scholar
  34. Rill RL, Hecker KH (1996) Sequence-specific actinomycin D binding to single-stranded DNA inhibits HIV reverse transcriptase and other polymerases. Biochemistry 35:3525–3533CrossRefPubMedGoogle Scholar
  35. Roos GHP, Loane C (2004) A clarifying NMR study of actinomycin X2. J Saudi Chem Soc 8:289–294Google Scholar
  36. Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425PubMedGoogle Scholar
  37. Shih HD, Liu YC, Hsu FL, Mulabagal V, Dodda R, Huang JW (2003) Fungichromin: a substance from Streptomyces padanus with inhibitory effects on Rhizoctonia solani. J Agric Food Chem 51:95–99CrossRefPubMedGoogle Scholar
  38. Shirling EB, Gottlieb D (1966) Methods for characterization of Streptomyces species. Int J Syst Bacteriol 16:313–340Google Scholar
  39. Shirling EB, Gottlieb D (1968) Cooperative description of type cultures of Streptomyces. III. Additional species descriptions from first and second studies. Int J Syst Bacteriol 18:279–392Google Scholar
  40. Takusagawa F, Carlson RG, Weaver RF (2001) Anti-leukemia selectivity in actinomycin analogues. Bioorg Med Chem 9:719–725CrossRefPubMedGoogle Scholar
  41. Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680PubMedCrossRefGoogle Scholar
  42. Wadkins RM, Vladu B, Tung CS (1998) Actinomycin D binds to metastable hairpins in single-stranded DNA. Biochemistry 37:11915–11923CrossRefPubMedGoogle Scholar
  43. Waksman SA, Woodruff HB (1940) Bacteriostatic and bacteriocidal substances produced by soil actinomycetes. Proc Soc Exp Biol Med 45:609–614Google Scholar
  44. Wayne LG, Brenner DJ, Colwell RR, Grimont PAD, Kandler O, Krichevsky MI, Moore LH, Moore WEC, Murray RGE, Stackebrandt E, Starr MP, Trüper HG (1987) Report of the Ad Hoc Committee on reconciliation of approaches to bacterial systematics. Int J Syst Bacteriol 37:463–464CrossRefGoogle Scholar
  45. Williams WK, Katz E (1977) Development of a chemically defined medium for the synthesis of actinomycin D by Streptomyces parvulus. Antimicrob Agents Chemother 11:281–290PubMedGoogle Scholar
  46. Womer RB (1997) Soft tissue sarcomas. Eur J Cancer 33:2230–2234; discussion 2234–2236CrossRefPubMedGoogle Scholar
  47. Zhou C, Wu Y, Dong L, Ye H, Wu J (1999) Studies on the mutation and fermentation of Hainanmycin producing strain. Zhongguo Kangshengsu Zazhi 24:11–13Google Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • K. Kurosawa
    • 1
  • V. P. Bui
    • 1
  • J. L. VanEssendelft
    • 1
  • L. B. Willis
    • 1
  • P. A. Lessard
    • 1
  • I. Ghiviriga
    • 2
  • T. G. Sambandan
    • 3
  • C. K. Rha
    • 3
  • A. J. Sinskey
    • 1
    • 4
  1. 1.Department of Biology, Building 68-370Massachusetts Institute of TechnologyCambridgeUSA
  2. 2.Department of ChemistryUniversity of FloridaGainesvilleUSA
  3. 3.Biomaterials Science and Engineering LaboratoryMassachusetts Institute of TechnologyCambridgeUSA
  4. 4.Department of Health Sciences and TechnologyMassachusetts Institute of TechnologyCambridgeUSA

Personalised recommendations