한국미생물·생명공학회지 (Microbiology and Biotechnology Letters)

  • 제38권2호
  • /
  • Pages.136-143
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  • 2010
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  • 1598-642X(pISSN)
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  • 2234-7305(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
권호 표지

국내에서 분리한 Sorangium cellulosum의 신규 Polyketide Synthase 유전자 검출

Detection of Novel Polyketide Synthase Genes in Sorangium cellulosum Isolated in Korea

  • 윤진권 (호서대학교 생명공학과 점액세균은행) ;
  • 김도희 (호서대학교 생명공학과 점액세균은행) ;
  • 이한빛 (호서대학교 생명공학과 점액세균은행) ;
  • 이계원 (호서대학교 생명공학과 점액세균은행) ;
  • 조경연 (호서대학교 생명공학과 점액세균은행)
  • Youn, Jin-Kwon (Myxobacteria Bank, Department of Biotechnology, Hoseo University) ;
  • Kim, Do-Hee (Myxobacteria Bank, Department of Biotechnology, Hoseo University) ;
  • Lee, Han-Bit (Myxobacteria Bank, Department of Biotechnology, Hoseo University) ;
  • Lee, Kye-Won (Myxobacteria Bank, Department of Biotechnology, Hoseo University) ;
  • Cho, Kyung-Yun (Myxobacteria Bank, Department of Biotechnology, Hoseo University)
  • 투고 : 2010.03.03
  • 심사 : 2010.05.17
  • 발행 : 2010.06.28
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초록

국내에서 분리한 9균주의 Sorangium cellulosum로부터 중합효소연쇄반응(PCR)을 통해 polyketide synthase(PKS)의 ketosynthase(KS) domain을 암호화하는 DNA를 증폭하고, 플라스미드 벡터에 클로닝한 후, 염기서열을 결정하였다. 전체 83개의 클로닝된 DNA 조각을 분석하여 유사한 조각을 배제한 결과, 43조각이 KS domain을 암호화하는 독립된 DNA 조각으로 판명되었다. 43조각 중 32조각의 아미노산 서열이 이미 클로닝된 PKS 유전자의 아미노산 서열과 70%-100% 유사하였으며, 나머지 11 조각은 알려진 서열과 67% 이하의 상동성을 가져 새로운 PKS 유전자일 가능성이 매우 높음을 보여주었다.

DNA fragments encoding the ketosynthase (KS) domain of polyketide synthase (PKS) genes were amplified using polymerase chain reaction (PCR) from 9 strains of Sorangium cellulosum isolated in Korea, cloned into a plasmid vector and sequenced. A total of 83 cloned DNA fragments were analyzed, and similar fragments were excluded, leaving 43 independent DNA fragments encoding the KS domains. The predicted amino acid sequences of 32 fragments were 70%-100% identical to the amino acid sequences of already known PKS genes, while the remaining 11 fragments were $\leq$67% or less identical to the known sequences, suggesting that these genes are novel PKS genes.

키워드

참고문헌

  1. Bode, H. B. and R. Müller. 2006. Analysis of myxobacterial secondary metabolism goes molecular. J. Ind. Microbiol. Biotechnol. 33: 577-588. https://doi.org/10.1007/s10295-006-0082-7
  2. Cane, D. E., C. T. Walsh, and C. Khosla. 1998. Harnessing the biosynthetic code: combinations, permutations, and mutations. Science 282: 63-68. https://doi.org/10.1126/science.282.5386.63
  3. Finking, R. and M. A. Marahiel. 2004. Biosynthesis of nonribosomal peptides. Annu. Rev. Microbiol. 58: 453-488. https://doi.org/10.1146/annurev.micro.58.030603.123615
  4. Frank, B., J. Knauber, H. Steinmetz, M. Scharfe, H. Blocker, S. Beyer, and R. Müller. 2007. Spiroketal polyketide formation in Sorangium: identification and analysis of the biosynthetic gene cluster for the highly cytotoxic spirangienes. Chem. Biol. 14: 221-233. https://doi.org/10.1016/j.chembiol.2006.11.013
  5. Gerth, K., O. Perlova, and R. Muller. 2008. Sorangium cellulosum. pp. 329-348. In Whitworth, D. E. (ed.), Myxobacteria: multicellularity and differentiation. ASM Press, Washington, D.C.
  6. Gerth, K., S. Pradella, O. Perlova, S. Beyer, and R. Muller. 2003. Myxobacteria: proficient producers of novel natural products with various biological activities-past and future biotechnological aspects with the focus on the genus Sorangium. J. Biotech. 106: 233-253. https://doi.org/10.1016/j.jbiotec.2003.07.015
  7. Hyun, H., J. Chung, H. Lee, J. Youn, C. Lee, D. Kim, and K. Cho. 2009. Isolation of cellulose-degrading myxobacteria Sorangium cellulosum. Kor. J. Microbiol. 45: 48-53.
  8. Hyun, H., J. Chung, J. Kim, J. S. Lee, B. -M. Kwon, K. -H. Son, and K. Cho. 2008. Isolation of Sorangium cellulosum carrying epothilone gene clusters. J. Microbiol. Biotechnol. 18: 1416-1422.
  9. Johnson, M., I. Zaretskaya, Y. Raytselis, Y. Merezhuk, S. McGinnis, and T. L. Madden. 2008. NCBI BLAST: a better web interface. Nucleic Acids Res. 36: W5-W9. https://doi.org/10.1093/nar/gkn201
  10. Julien, B., S. Shah, R. Ziermann, R. Goldman, L. Katz, and C. Khosla. 2000. Isolation and characterization of the epothilone biosynthetic gene cluster from Sorangium cellulosum. Gene 16: 153-160.
  11. Julien, B., Z. Q. Tian, R. Reid, and C. D. Reeves. 2006. Analysis of the ambruticin and jerangolid gene clusters of Sorangium cellulosum reveals unusual mechanisms of polyketide biosynthesis. Chem. Biol. 13: 1277-1286. https://doi.org/10.1016/j.chembiol.2006.10.004
  12. Kim, Y. S., W. C. Bae, and S. J. Back. 2003. Bioactive substances from myxobacteria. Kor. J. Microbiol. Biotechnol. 31: 1-12.
  13. Komaki, H., R. Fudou, T. Iizuka, D. Nakajima, K. Okazaki, D. Shibata, M. Ojika, and S. Harayama. 2008. PCR detection of type I polyketide synthase genes in myxobacteria. Appl. Environ. Microbiol. 74: 5571-5574. https://doi.org/10.1128/AEM.00224-08
  14. Larkin, M. A., G. Blackshields, N. P. Brown, R. Chenna, P. A. McGettigan, H. McWilliam, F. Valentin, I. M. Wallace, A. Wilm, R. Lopez, J. D. Thompson, T. J. Gibson, and D. G. Higgins. 2007. Clustal W and Clustal X version 2.0. Bioinformatics 23: 2947-2948. https://doi.org/10.1093/bioinformatics/btm404
  15. Li, Z. F., J. Y. Zhao, Z. J. Xia, J. Shi, H. Liu, Z. H. Wu, W. Hu, W. F. Liu, and Y. Z. Li. 2007. Evolutionary diversity of ketoacyl synthases in cellulolytic myxobacterium Sorangium. Syst. Appl. Microbiol. 30: 189-196. https://doi.org/10.1016/j.syapm.2006.06.002
  16. Molnar, I., T. Schupp, M. Ono, R. Zirkle, M. Milnamow, B. Nowak-Thompson, N. Engel, C. Toupet, A. Stratmann, D. D. Cyr, J. Gorlach, J. M. Mayo, A. Hu, S. Goff, J. Schmid, and J. M. Ligon. 2000. The biosynthetic gene cluster for the microtubule-stabilizing agents epothilones A and B from Sorangium cellulosum So ce90. Chem. Biol. 7: 97-109. https://doi.org/10.1016/S1074-5521(00)00075-2
  17. Reichenbach, H. 2005. Myxococcales. pp. 1059-1144. In Brenner, D.J., N.R. Krieg, J.T. Staley, and G.M. Garrity (ed.), Bergey's Manual of Systematic Bacteriology, 2nd ed. Bergey's Manual Trust, East Lansing, MI.
  18. Reichenbach, H. and M. Dworkin. 1992. The myxobacteria. pp. 3416-3487. In Balows, A., H.G. Trüper, M. Dworkin, W. Harder, and K.-H. Schleifer (ed.), The Prokaryotes, 2nd ed., vol. IV, Springer Verlag, NY.
  19. Ronning, C. M. and W. C. Nierman. 2008. The genomes of Myxococcus xanthus and Stigmatella aurantiaca, pp. 285-298. In Whitworth, D. E. (ed.), Myxobacteria: multicellularity and differentiation. ASM Press, Washington, D.C.
  20. Sambrook, J., E. F. Fritsch and T. Maniatis. 1989. Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.
  21. Schirmer, A., R. Gadkari, C. D. Reeves, F. Ibrahim, E. F. DeLong, and C. R. Hutchinson. 2005. Metagenomic analysis reveals diverse polyketide synthase gene clusters in microorganisms associated with the marine sponge Discodermia dissoluta. Appl. Environ. Microbiol. 71: 4840-4849. https://doi.org/10.1128/AEM.71.8.4840-4849.2005
  22. Schneiker, S., O. Perlova, O. Kaiser, K. Gerth, A. Alici, M. O. Altmeyer, D. Bartels, T. Bekel, S. Beyer, E. Bode, H. B. Bode, C. J. Bolten, J. V. Choudhuri, S. Doss, Y. A. Elnakady, B. Frank, L. Gaigalat, A. Goesmann, C. Groeger, F. Gross, L. Jelsbak, L. Jelsbak, J. Kalinowski, C. Kegler, T. Knauber, S. Konietzny, M. Kopp, L. Krause, D. Krug, B. Linke, T. Mahmud, R. Martinez-Arias, A. C. McHardy, M. Merai, F. Meyer, S. Mormann, J. Munoz-Dorado, J. Perez, S. Pradella, S. Rachid, G. Raddatz, F. Rosenau, C. Ruckert, F. Sasse, M. Scharfe, S. C. Schuster, G. Suen, A. Treuner-Lange, G. J. Velicer, F.J. Vorholter, K. J. Weissman, R. D. Welch, S. C. Wenzel, D. E. Whitworth, S. Wilhelm, C. Wittmann, H. Blocker, A. Puhler, and R. Muller. 2007. Complete genome sequence of the myxobacterium Sorangium cellulosum. Nat. Biotechnol. 25: 1281-1289. https://doi.org/10.1038/nbt1354
  23. Staunton, J. and B. Wilkinson. 2001. Combinatorial biosynthesis of polyketides and nonribosomal peptides. Curr. Opin. Chem. Biol. 5: 159-164. https://doi.org/10.1016/S1367-5931(00)00185-X
  24. Staunton, J. and K. J. Weissman. 2001. Polyketide biosynthesis: a millennium review. Nat. Prod. Rep. 18: 380-416. https://doi.org/10.1039/a909079g
  25. Weissman, K. J. and R. Müller. 2009. A brief tour of myxobacterial secondary metabolism. Bioorg. Med. Chem. 17: 2121-2136. https://doi.org/10.1016/j.bmc.2008.11.025