Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지

Isolation and Characterization of the Eicosapentaenoic Acid Biosynthesis Gene Cluster from Shewanella sp. BR-2

  • Lee, Su-Jin (Molecular Bioprocess Research Center, Jeonbuk Branch Institute, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Seo, Pil-Soo (Molecular Bioprocess Research Center, Jeonbuk Branch Institute, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, Chul-Ho (Molecular Bioprocess Research Center, Jeonbuk Branch Institute, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kwon, Oh-Suk (Omics and Integration Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Hur, Byung-Ki (Department of Biological Engineering and Institute of Biotechnological Industry, Inha University) ;
  • Seo, Jeong-Woo (Molecular Bioprocess Research Center, Jeonbuk Branch Institute, Korea Research Institute of Bioscience and Biotechnology (KRIBB))
  • Published : 2009.09.30
Download PDF
⟨ Previous Next ⟩

Abstract

Forty-four eicosapentaenoic acid (EPA)-producing microbial strains were isolated from the intestines of marine fishes. Among them, one strain showing a maximum level of EPA (4.78% of total fatty acids) was identified as Shewanella sp. BR-2 on the basis of its 168 rRNA sequence. The EPA content reached a maximum level during the mid-exponential phase of cell growth, and gradually decreased with further growth of the cells. A cosmid DNA including the EPA biosynthesis gene cluster consisting of pfaA-E was isolated from a cosmid library of genomic DNA of Shewanella sp. BR-2, named pCosEPA-BR2. An E. coli clone harboring pCosEPA-BR2 produced EPA at a maximum level of 7.5% of total fatty acids, confirming the EPA biosynthesis activity of the cloned gene cluster.

Keywords

References

  1. Abbey, M., P. Clifton, M. Kestin, B. Belling, and P. Nestel. 1990. Effect of fish oil on lipoproteins, lecithin:cholesterol acyltransferase, and lipid transfer protein activity in humans. Arteriosclerosis 10: 85-94 https://doi.org/10.1161/01.ATV.10.1.85
  2. Angerer, P. and C. V. Schachy. 2000. n-3 Polyunsaturated fatty acids and the cardiovascular system. Curr. Opin. Clin. Nutr. Metab. Care 3: 439-445 https://doi.org/10.1097/00075197-200011000-00005
  3. Braden, L. M. and K. K. Caroll. 1986. Dietary polyunsaurated fat in relation to mammary carcinogenesis in rats. Lipids 21: 285-288 https://doi.org/10.1007/BF02536414
  4. Chung, T. H., S. J. Lee, H. J. Oh, G. J. Kim, and B. K. Hur. 2005. Cloning of a $\Delta$5 desaturase from Thraustochytrium sp. 26185 and functional expression in Pichia pastoris. Biotechnol. Bioprocess Eng. 20: 93-99
  5. Fiocchi, A., M. Sala, P. Signoroni, G. Banderal, C. Agostoul, and E. Riva. 1994. The efficiency and safety of $\gamma$-linolenic acid in the treatment of infantile atopic dermatitis. J. Int. Med. Res. 22: 2-32
  6. Jiang, H., R. Zirkle, J. G. Metz, L. Braun, L. Richter, S. G. Van Lanen, and B. Shen. 2008. The role of tandem acyl carrier protein domains in polyunsaturated fatty acid biosynthesis. J. Am. Chem. Soc. 130: 6336-6337 https://doi.org/10.1021/ja801911t
  7. Kaulmann, U. and C. Hertweck. 2002. Biosynthesis of polyunsaturated fatty acids by polyketide synthases. Angew. Chem. Int. Ed. 41: 1866-1869 https://doi.org/10.1002/1521-3773(20020603)41:11<1866::AID-ANIE1866>3.0.CO;2-3
  8. Kimura, M. 1980. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J. Mol. Evol. 16: 111-120 https://doi.org/10.1007/BF01731581
  9. Lauritzen, L., H. S. Hansen, M. H. Jorgensen, and K. F. Michaelsen. 2001. The essentiality of long chain n-3 fatty acids in relation to development and function of the brain and retina. Prog. Lipid Res. 40: 1-94 https://doi.org/10.1016/S0163-7827(00)00017-5
  10. Lee, S. J., Y. S. Jeong, D. U. Kim, J.-W. Seo, and B. K. Hur. 2006. Eicosapentaenoic acid (EPA) biosynthetic gene cluster of Shewanella oneidensis MR-1: Cloning, heterologous expression, and effect of temperature and glucose on the production of EPA in Escherichia coli. Biotech. Bioprocess Eng. 11: 510-515 https://doi.org/10.1007/BF02932075
  11. Medina, A. R., E. M. Grima, A. G. Gimenez, and M. J. I. Gonzalez. 1998. Downstream processing of algal polyunsaturated fatty acids. Biotechnol. Adv. 16: 517-580 https://doi.org/10.1016/S0734-9750(97)00083-9
  12. Metz, J. G., P. Roessler, D. Facciotti, C. Levering, F. Dittrich, M. Lassner, et al. 2001. Production of polyunsaturated fatty acids by polyketide synthases in both prokaryotes and eukaryotes. Science 293: 290-293 https://doi.org/10.1126/science.1059593
  13. Okuyama, H., Y. Orikasa, T. Nishida, K. Watanabe, and N. Norita. 2007. Bacterial genes responsible for the biosynthesis of eicosapentaenoic and docosaphexenoic acids and their heterologous expression. Appl. Environ. Microbiol. 73: 665-670 https://doi.org/10.1128/AEM.02270-06
  14. Orikasa, Y., A. Yamada, R. Yu, Y. Ito, T. Nishida, I. Yumoto, K. Watanabe, and H. Okuyama. 2004. Characterization of the eicosapentaenoic acid biosynthesis gene cluster from Shewanella sp. strain SCRC-2738. Cell Mol. Biol. 50: 625-630
  15. Qiu, X. 2003. Biosynthesis of docosahexaenoic acid (DHA, 22:6-4, 7,10,13,16,19): Two distinct pathways. Prostaglandins Leukot. Essen. Fatty Acids 68: 181-186 https://doi.org/10.1016/S0952-3278(02)00268-5
  16. Sauer, L. A., R. T. Dauchy, and D. E. Blask. 2001. Polyunsaturated fatty acids, melatonin, and cancer prevention. Biochem. Pharmacol. 61: 1455-1462 https://doi.org/10.1016/S0006-2952(01)00634-7
  17. Tamura, K., J. Dudley, M. Nei, and S. Kumar. 2007. MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol. Biol. Evol. 24:1596-1599 https://doi.org/10.1093/molbev/msm092
  18. 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
  19. Trautwein, E. A. 2001. n-3 Fatty acids - physiological and technical aspects for their role in food. Eur. J. Lipid Sci. Technol. 103: 45-55 https://doi.org/10.1002/1438-9312(200101)103:1<45::AID-EJLT45>3.0.CO;2-9
  20. Tresguerres, J. A. F., V. Lahera, and M. V. Cachofeiro. 1989. Dietary essential polyunsaturated fatty acids and blood pressure. News Physiol. Sci. 4: 64-68
  21. Wallis, J. G., J. L. Watts, and J. Browse. 2002. Polyunsaturated fatty acid synthesis: What will they think of next? Trends Biochem. Sci. 27: 467-473 https://doi.org/10.1016/S0968-0004(02)02168-0
  22. Yazawa, K. 1996. Production of eicosapentaenoic acid from marine bacteria. Lipids 31: S297-S300 https://doi.org/10.1007/BF02637095