생명과학회지 (Journal of Life Science)

  • 제19권11호
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  • Pages.1522-1528
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  • 2009
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
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해양미생물 Streptomyces sp. M3로부터 alginate lyase의 클로닝 및 발현

Cloning and Expression of Alginate Lyase from a Marine Bacterium, Streptomyces sp. M3

  • 김희숙 (경성대학교 식품생명공학과)
  • Kim, Hee-Sook (Department of Food Science and Biotechnology, Kyungsung University)
  • 발행 : 2009.11.30
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초록

알긴산을 분해하기 위하여 갈조류로부터 분해활성이 있는 해양미생물을 분리하였다. 분리된 균주의 16S ribosomal DNA를 분석한 결과 이전에 보고했던 ALG-5 균주와 비슷한 Streptomyces sp.에 속하는 것으로 나타났다. 상동성이 있는 염기서열로 고안한 특이적인 primer로 PCR을 행함로서 Streptomyces sp. M3의 새로운 alginate lyase 유전자를 클로닝하였다. M3 alginate lyase의 예상 아미노산 서열에는 N-terminal 영역에 YXRSELREM 서열과 C-terimnal 영역에 YFKAGXYXQ 서열이 보존되어 있었다. M3 alginate lyase 단백질의 homology model은 Corynebacterium sp. ALY-1으로부터 얻은 단백질인 alyPG와 같이 $\beta$-jelly roll fold를 main domain으로 가지고 있음이 나타났다. M3 alginate lyase 유전자를 가지는 재조합 E. coli의 세포균질액은 polymannuronate block보다는 polyguluronate block에 대하여 높은 분해력을 가지고 있었다. 아미노산 서열 다중정열 및 homology modeling으로부터 얻은 결과는 M3 alginate lyase가 Family PL-7으로 분류될 수 있음을 말해 준다.

A marine bacterium was isolated from brown seaweeds for its ability to degrade alginate. Analysis of 16S ribosomal DNA sequence revealed that the strain belongs to Streptomyces like strain ALG-5 which was reported previously. New alginate lyase gene of Streptomyces sp. M3 was cloned by using PCR with the specific primers designed from homologous nucleotide sequences. The consensus sequences of N-terminal YXRSELREM and C-terminal YFKAGXYXQ were conserved in the M3 alginate lyase amino acid sequences. The homology model for the M3 alginate lyase showed a characteristic structure of $\beta$-jelly roll fold main domain like alyPG from Corynebacterium sp. ALY-1. The homogenate of the recombinant E. coli with the alginate lyase gene showed more degrading activity for polyguluronate block than polymannuronate block. The results from the multiple alignments and the homology modeling elucidated in the M3 alginate lyase can be classified into family PL-7.

키워드

참고문헌

  1. Akiyama, H., T. Endo, R. Nakakita, K. Murata, Y. Yonemoto, and K. Okayama. 1992. Effect of depolymerized alginates on the growth of bifidobacteria. Biosci. Biotechnol. Biochem. 56, 355-356 https://doi.org/10.1271/bbb.56.355
  2. Cao, L., L. Xie, X. Xue, H. Tan, Y. Liu, and S. Zhou. 2007. Purification and characterization of alginate lyase from Streptomyces species strain A5 isolated from banana rhizosphere. J. Agric. Food. Chem. 55, 5113-5117 https://doi.org/10.1021/jf0704514
  3. Choi, S. H., H. S. Kim, and E. Y. Lee. 2008. Enantioselective kinetic resolution of racemic styrene oxide using recombinant marine fish epoxide hydrolase of Mugil cephalus. J. Korean Ind. Eng. Chem. 19, 491-496
  4. Cote, G. L. and L. H. Krull. 1988. Characterization of the extracellular polysaccharides from Azotobacter chroococcum. Carbohydr. Res. 181, 143-152 https://doi.org/10.1016/0008-6215(88)84030-8
  5. Fan, M. Y., Z. P. Lum, X. W. Fu, L. Levesque, I. T. Tai, and A. M. Sun. 1990. Reversal of diabetes in BB rats by transplantation of encapsulated pancreatic islets. Diabetes 39, 519-522 https://doi.org/10.2337/diabetes.39.4.519
  6. Grovan, J. R. W., J. A. M. Fyfe, and T. R. Jarman. 1981. Isolation of alginate-producing mutants of Pseudomonas fluorescens, Pseudomonas putida and Pseudomonas mendocina. J. Gen. Microbiol. 125, 217-220 https://doi.org/10.1099/00221287
  7. Haug, A., B. Larsen, and O. Smidsrot. 1966. A study of the constitution of alginic acid by partial acid hydrolysis. Acta. Chem. Scand. 20, 183-190 https://doi.org/10.3891/acta.chem.scand.20-0183
  8. Iwamoto, M., M. Kurachi, T. Nakashima, D. Kim, K. YamaguchI, T. Oda, Y. Iwamoto, and T. Muramatsu. 2005. Structure-activity relationship of alginate oligosaccharides in the induction of cytokine production from RAW264.7 cells. FEBS Lett. 579, 4423-4429 https://doi.org/10.1016/j.febslet.2005.07.007
  9. Kawada, A., N. Hiura, S. Tajima, and H. Takahara. 1999. Alginate oligosaccharides stimulate VEGF-mediated growth and migration of human endothelial cells. Arch. Dermatol. Res. 291, 542-547 https://doi.org/10.1007/s004030050451
  10. Kawamoto, H., A. Horibe, Y. Miki, T. Kimura, K. Tanaka, T. Nakagawa, M. Kawamukai, and H. Matsuda. 2006.Cloning and sequencing analysis of alginate lyase genes from the marine bacterium Vibrio sp. 02. Mar. Biotechnol. 8, 481-490 https://doi.org/10.1007/s10126-005-6157-z
  11. Kim, D. E., E. Y. Lee, and H. S. Kim. 2009. Cloning and characterization of alginate lyase from a marine bacterium Streptomyces sp. ALG-5. Mar. Biotechnol. 11, 10-16 https://doi.org/10.1007/s10126-008-9114-9
  12. Kim, H. S., O. K. Lee, S. J. Lee, S. Hwang, S. J. Kim, S. Yang, S. Park, and E. Y. Lee. 2006. Enantioselective epoxide hydrolase activity of a newly isolated microorganism, Sphingomonas echinoids EH-983, from seawater. J. Mol. Catal. B: Enzym. 41, 130-135 https://doi.org/10.1016/j.molcatb.2006.05.009
  13. Matsubara, Y., R. Kawada, K. Iwasaki, Y. Kimura, T. Oda, and T. Muramatsu. 2000. Cloning and sequence analysis of a gene (aly PG) encoding poly(a-L-guluronate) lyase from Corynebacterium sp. Strain ALY-1. J. Biosci. Bioeng. 89, 199-202 https://doi.org/10.1016/S1389-1723(00)88738-1
  14. Osawa, T., Y. Matsubara, T. Muramatsu, M. Kimura, and Y. Kakuta. 2005. Crystal structure of the alginate (poly-$\alpha$-Lguluronate) lyase from Corynebacterium sp. at 1.2 ${\AA}$ resolution. J. Mol. BioI. 345, 1111-1118 https://doi.org/10.1016/j.jmb.2004.10.081
  15. Quing, G., L.-C. Ma, A. Khorchid, G. V. T. Swapna, T. K. Mal,l M. M. Takayama, B. Xia, S. Phadtare, H. Ke, T. Acton, G. T. Montelione, M. Ikura, and M. Inouye. 2004. Cold-shock induced high-yield protein production in Escherichia coli. Nature Biotech. 22, 877-882 https://doi.org/10.1038/nbt984
  16. Schirano, Y. and D. Shibata. 1990. Low temperature cultivation of Escherichia coli carrying a rice lip oxygenase L-2 cDNA produces a soluble and active enzyme at a high level. FEBS Lett. 271. 128-130 https://doi.org/10.1016/0014-5793(90)80388-Y
  17. Scott, C. D. 1987. Immobilized cells: a review of recent literature. Enzyme Microb. Technol. 9, 66-73 https://doi.org/10.1016/0141-0229(87)90145-1
  18. Wong, T. Y., L. A. Preston, and N. L. Schiller. 2000. Alginate lyase: Review of major sources and enzyme characteristics, structure-function analysis, biological roles, and application. Annu. Rev. Microbiol. 54, 289-340 https://doi.org/10.1146/annurev.micro.54.1.289
  19. Yamasaki, M., K. Ogura, W. Hashimoto, B. Mikami, and K. Murata. 2005. A structural basis for depolymerization of alginate by polysaccharide lyase family-7. J. Mol. BioI. 352, 11-21 https://doi.org/10.1016/j.jmb.2005.06.075
  20. Yamasaki, M., S. Moriwaki, O. Miyake, W. Hashimoto, K. Murata, and B. Mikami. 2004. Structure and function of a hypothetical Psedomonas aeruginosa protein PA1167 classified into family PL-7. J. BioI. Chem. 279, 31863-31872 https://doi.org/10.1074/jbc.M402466200
  21. Yanemoto, Y., H. Tanaka, T. Yamashita, N. Kitabatake, Y. Ishida, A. Kimura, and K. Murata. 1993. Promotion of germination and shoot elongation of some plants by alginate oligomers prepared with bacterial alginate lyase. Ferment. Bioeng. 75, 68-70 https://doi.org/10.1016/0922-338X(93)90181-7
  22. Yoon, H., W. Hashimoto, O. Miyake, M. Okamoto, B. Mikami, and K. Murata. 2000. Overexpression in Escherichia coli, purification, and characterization of Sphingomonas sp. Al alginate lyase. Protein Expres. Purif. 19, 84-90 https://doi.org/10.1006/prep.2000.1226

피인용 문헌

  1. Characterization of Recombinant PolyG-Specific Lyase from a Marine Bacterium, Streptomyces sp. M3 vol.20, pp.11, 2010, https://doi.org/10.5352/JLS.2010.20.11.1582
  2. Enhancing the Alginate Degrading Activity of Streptomyces sp. Strain M3 Alginate Lyase by Mutation vol.22, pp.1, 2012, https://doi.org/10.5352/JLS.2012.22.1.7
  3. Biochemical Characterization and Degradation Pattern of a Novel Endo-Type Bifunctional Alginate Lyase AlyA from Marine Bacterium Isoptericola halotolerans vol.16, pp.8, 2018, https://doi.org/10.3390/md16080258