Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Isolation of Myxobacteria Carrying Soraphen Biosynthetic Gene Clusters

Soraphen 생합성 유전자군을 갖는 점액세균의 분리

  • Lee, Cha-Yul (Myxobacteria Bank, Department of Biotechnology, Hoseo University) ;
  • Hyun, Hye-Sook (Myxobacteria Bank, Department of Biotechnology, Hoseo University) ;
  • Cho, Kung-Yun (Myxobacteria Bank, Department of Biotechnology, Hoseo University)
  • 이차율 (호서대학교 생명공학과 점액세균은행) ;
  • 현혜숙 (호서대학교 생명공학과 점액세균은행) ;
  • 조경연 (호서대학교 생명공학과 점액세균은행)
  • Published : 2009.03.28
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Abstract

Polymerase chain reactin (PCR) could be a simple way to screen new microbial strains producing useful secondary metabolites if their biosynthetic genes are known and candidate strains to be screened are available. In this study, we have screened two myxobacterial strains, KYC3047 and KYC3076, carrying genes appeared to be biosynthetic genes of soraphen A, a potent antifungal substance, out of 50 cellulose degrading myxobacteria using PCR. The two strains were identified as Sorangium cellulosum based on morphological, physiological, and molecular biological characteristics. Both of the strains produced substances having strong antifungal activities as expected against Candida albicans, a causative agent of candidiasis, and Colletotrichum acutatum, a causative agent of anthracnose on pepper.

생합성 유전자들이 알려져 있고 탐색할 후보 균주들이 확보되어있는 경우에 중합효소연쇄반응(PCR)은 유용한 이차 대사산물을 생산하는 새로운 균주의 탐색에 있어서 간편한 방법이 될 수 있다. 본 연구에서는 중합효소연쇄반응에 의해 50균주의 셀룰로오스 분해성 점액세균로부터 강력한 항진균물질 soraphen의 생합성 유전자로 보이는 유전자를 가지는 두 점액세균 균주 KYC3047과 KYC3076를 선별하였다. 그리고 형태학적, 생리학적, 분자생물학적 특성에 의해 이들 균주들을 S. cellulosum로 동정하였다. 두 균주는 모두 예상한대로 칸디다증을 유발하는 Candida albicans와 고추 탄저병을 유발하는 Colletotrichum acutatum에 강한 항균활성을 갖는 물질을 생산하였다.

Keywords

References

  1. Beckers, A., S. Organe, L. Timmermans, K. Scheys, A. Peeters, K. Brusselmans, G. Verhoeven, and J. V. Swinnen. 2007. Chemical inhibition of acetyl-CoA carboxylase induces growth arrest and cytotoxicity selectively in cancer cells. Cancer Res. 67: 8180-8187 https://doi.org/10.1158/0008-5472.CAN-07-0389
  2. Chung, J., C. Lee, S.-C. Yun, and K. Cho. 2008. Screening of myxobacteria inhibiting the growth of Collectotrichum acutatum causing anthracnose on pepper. Kor. J. Microbiol. Biotechnol. 36: 21-27 (in Korean with English abstract)
  3. Gerth, K., N. Bedorf, H. Irschik, G. Hfle, and H. Reichenbach. 1994. The soraphens: a farni1y of novel anti fungal compounds from Sorangium cellulosum (Myxobacteria). J. Antibiot. 47: 23-31
  4. Gerth, K., S. Pradella, O. Perlova, S. Beyer, and R. Mller. 2003. Myxobacteria: proficient producers of novel natural products with various biologica1 activities - past and future biotechnological aspects with the focus on the genus Sorangium. J. Biotechnol. 106: 233-253 https://doi.org/10.1016/j.jbiotec.2003.07.015
  5. Hill, A. M. and B. L. Thompson. 2003. NoveL soraphens from precursor directed biosynthesis. Chem. Commun. 21: 1360-1361
  6. Kim, Y. S., W. C. Bae, and S. J. Back. 2003. Bioactive substances from myxobacteria. Kor. J. Microbiol. Biotechnol. 31: 1-12 (in Korean with English abstract) https://doi.org/10.1007/s10295-004-0109-x
  7. Lee, H. W., I.-Y. Choi Lee, Y.-S. Kim, and S.-U. Park. 2002 An efficient synthesis of C3-C9 segment of soraphen A. Bull. Korean Chem. Soc. 23: 1197-1198
  8. Ligon J, S. Hill, J. Beck, R. Zirkle, I. Mo1nr, J. Zawodny, S. Money, and T. Schupp. 2002. Characterization of the biosynthetic gene cluster for the antifungal polyketide soraphen A from Sorangium cellulosum So ce26. Gene 285: 257-267 https://doi.org/10.1016/S0378-1119(02)00396-7
  9. Park, S. H. and H. W. Lee. 2008. Synthetic studies on soraphen A: synthesis of the C1-C9 hemiketal segment. Bull. Korean Chem. Soc. 29: 1445-1446 https://doi.org/10.5012/bkcs.2008.29.8.1445
  10. Park, S. H., H. W. Lee, S.-U. Park. 2004. Synthesis of C3-C9 suLfonyL derivative of soraphen A. Bull. Korean Chem. Soc. 25: 1613-1614 https://doi.org/10.5012/bkcs.2004.25.11.1613
  11. Rainey, F. A., Rainey, W. N., Kroppenstedt, R. M., and Stackerbrandt, E. 1996. The Genus Nocardiopsis represents a phylogenetically coherent taxon and a distinct actinomycete lineage: proposal of Nocardiopsaceae fam. nov. lnt. J. Syst. Bacteriol. 46: 1088-1092 https://doi.org/10.1099/00207713-46-4-1088
  12. Reichenbach, H. 2005. Myxococcales. pp. 1059-1144. ln Brenner, D. J., N. R. Krieg, J. T. Staley, and G. M. Garrity(eds.), Bergey's Manual of Systematic Bacteriology, 2nd ed. Bergey's Manual Trust, East Lansing, MI
  13. Reichenbach, H. and M. Dworkin. 1992. The myxobacteria. pp. 3416-3487. ln Balows, A., H. G Trper, M. Dworkin, W. Harder, and K. H. Schleifer (eds.), The Prokaryotes, 2nd ed. vol. IV,Springer Verlag, New York
  14. Schupp, T. C. Toupet, B. Cluzel, S. Neff, S. Hill, J. J. Beck, J. M. Ligon. 1995. A Sorangium cellulosum (myxobacterium) gene cluster for the biosynthesis of the macrolide antibiotic soraphen A:cloning, characterization, and homology to poiyketide synthase genes from actinomycetes. J. Bacteriol. 177: 3673-3679
  15. Shen, Y., S. L. Volrath, S. C. Weatherly, T. D. Elich, and L. Tong. 2004. A mechanism for the potent inhibition of eukaryotic acetyl-coenzyme A carboxylase by soraphen A, a macrocyclic polyketide natural product. Mol. Cell. 16: 881-891 https://doi.org/10.1016/j.molcel.2004.11.034
  16. Sprer, C., H. Reichenbach, and E. Stackebrandt. 1999. The correlation between morphological and phylogenetic classification of myxobacteria. Int. J. Syst. Bacteriol. 3: 1255-1262 https://doi.org/10.1099/00207713-49-3-1255
  17. Thompson, J. D., D. G. Higgins, and T. J. Gibson. 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: 4673-4680 https://doi.org/10.1093/nar/22.22.4673
  18. Vahlensieck, H. F., L. Pridzun, H. Reichenbach, and A. Hinnen. 1994. Identification of the yeast ACCl gene product (acetyl-CoA carboxylase) as the target of the polyketide fungicide soraphen A. Curr. Genet. 25: 95-100 https://doi.org/10.1007/BF00309532
  19. Zhang, H., B. Tweel, J. Li, and L. Tong, L. 2004. Crystal structure of the carboxyltransferase domain of acetylcoenzyme A carboxylase in complex with CP-640186. Structure 12: 1683-1691 https://doi.org/10.1016/j.str.2004.07.009
  20. Zhang, H., Z. Yang, Y. Shen, and L. Tong. 2003, Crystal structure of the carboxyltransferase domain of acetylcoenzyme A carboxylase. Science 299: 2064-2067 https://doi.org/10.1126/science.1081366
  21. Zirkle, R., J. M. Ligon, and I. Molnr I. 2004. Heterologous production of the antifungal polyketide antibiotic soraphen A of Sorangium cellulosum So ce26 in Streptomyces Itvidans. Microbiology 150: 2761-2774 https://doi.org/10.1099/mic.0.27138-0