Fisheries and Aquatic Sciences

The Korean Society of Fisheries and Aquatic Science (한국수산과학회)
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Characterization of Geranylgeranyl Pyrophosphate Synthase from the Marine Bacterium, Paracoccus haeundaensis

  • Seo, Yong-Bae (Department of Microbiology, Pukyong National University) ;
  • Lee, Jae-Hyung (Basic Science Research Institute, Pukyong National University) ;
  • Kim, Young-Tae (Department of Microbiology, Pukyong National University)
  • Published : 2009.03.31
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Abstract

Carotenoids such as $\beta$-carotene and astaxanthin are used as food colorants, animal feed supplements and for nutritional and cosmetic purposes. In a previous study, an astaxanthin biosynthesis gene cluster was isolated from the marine bacterium, Paracoccus haeundaensis. Geranylgeranyl pyrophosphate (GGPP) synthase (CrtE), encoded by the ortE gene, catalyzes the formation of GGPP from farnesyl pyrophosphate (FPP), which is an essential enzyme for the biosynthesis of carotenoids in early steps. In order to study the biochemical and enzymatic characteristics of this important enzyme, a large quantity of purified GGPP synthase is required. To overproduce GGPP synthase, the crtE gene was subcloned into a pET-44a(+) expression vector and transformed into the Escherichia coli BL21(DE3) codon plus cell. Transformants harboring the crtE gene were cultured and the crtE gene was over-expressed. The expressed protein was purified to homogeneity by affinity chromatography and applied to study its biochemical properties and molecular characteristics.

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References

  1. Akyon, Y. 2002. Effect of antioxidants on the immune response of Helicobacter pylori. Clin. Microbiol. Infect., 8, 438-441 https://doi.org/10.1046/j.1469-0691.2002.00426.x
  2. Amar, Y., V. Kiron, S. Satoh and T. Watanabe. 2004. Enhancemεnt of innate immunity in rainbow trout (Oncorhynchus mykiss Walbaum) associated with dietary intake of carotenoids from natural products. Fish Shellfish Immunol., 16, 527-537 https://doi.org/10.1016/j.fsi.2003.09.004
  3. Bertra, J. S. 1999. Carotenoids and gene regulation. Nutrition Reviews, 57, 182-19 1 https://doi.org/10.1111/j.1753-4887.1999.tb06941
  4. Chemler, J.A., Y. Yan and M.A. Koffas. 2006. Biosynthesis of isoprenoids, polyunsaturatεd fatty acids and flavonoids in Saccharomyces cerevisiae. Microb. Cell Fact., 23, 5-20
  5. Edge, R., D.J. McGarvey and T.G. Truscott. 1997. The carotεnoids as anti-oxidants a review. J. Photoch Photobio. B., 41, 189-200 https://doi.org/10.1016/S1011-1344(97)00092-4
  6. Jyonouchi, H., L. Ahang, M. Cross and Y. Tomita. 1994. lmunomodulating actions of carotenoids: enhancement of in vivo and in vitro antibody production to T -dependent antigens. Nutr. Cancer，21, 47-58 https://doi.org/10.1080/01635589409514303
  7. Lee, J.H. and Y.T. Kim. 2006. Cloning and characterization of the astaxanthin biosynthesis gene cluster from the marine bacterium Paracoccus haeundaensis. Gene, 370, 86-95 https://doi.org/10.1016/j.gene.2005.11.007
  8. Lee, J.H. and Y.T. Kim. 2006. Functional expression ofthe astaxanthin biosynthesis genes from a marine bacterìum, Paracoccus haeundaensis. Biotechnol. Lett., 28, 1167-1173 https://doi.org/10.1007/s10529-006-9072-0
  9. Lee，J.H., Y.S. Kim, T.J. Choi, W.J. Lee and Y.T. Kim. 2004. Paracoccus haeundaensis sp. nov., a Gramnegative, halophilic, astaxanthin-producing bacterium. lnt. J. Syst. Evol. Microbiol., 54, 1699-1702 https://doi.org/10.1099/ijs.0.63146-0
  10. Lee, J.H., Y.B. Seo, S.Y. Jeong, S.W. Nam and Y.T. Kim. 2007. Functional analysis of combinations in astaxanthin biosynthesis genes from Paracoccus haeundaensis. Biotecnol. Bioprocess Eng., 12, 312-317 https://doi.org/10.1007/BF02931110
  11. Math, S.K., J.E. Hearst and C.D. Poulter. 1992. The crtE gene in Erwinia herbicola encodes geranylgeranyldiphosphate synthase. Proc. Natl. Acad. Sci. USA, 89, 6761-6764 https://doi.org/10.1073/pnas.89.15.6761
  12. Wang, K. and S. Ohnuma. 1999. Chain-length determina-tion mechanism of isoprenyl diphosphate synthases and implications for molecular evolution. Trends Biochem. Sci., 24, 445-451 https://doi.org/10.1016/S0968-0004(99)01464-4