Characterization of Geranylgeranyl Pyrophosphate Synthase from the Marine Bacterium, Paracoccus haeundaensis
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Carotenoids such as β-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.
Abstract
Carotenoids such as β-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.
참고문헌 (12)
- Akyon, Y, "Effect of antioxidants on the immune response of Helicobacter pylori", Clin. Microbiol. Infect., 8, pp.438~441 (2002)
- Amar, Y., "Enhancement of innate immunity in rainbow trout (Oncorhynchus mykiss Walbaum) associated with dietary intake of carotenoids from natural products", Fish Shellfish Immunol., 16, pp.527~537 (2004)
- Bertra, J.S, "Carotenoids and gene regulation", Nutrition Reviews, 57, pp.182~191 (1999)
- Chemler, J.A., "Bio-synthesis of isoprenoids, polyunsaturated fatty acids and flavonoids in Saccharomyces cerevisiae", Microb. Cell Fact., 23, pp.5~20 (2006)
- Edge, R., "The carotenoids as anti-oxidants a review", J. Photoch. Photobio. B., 41, pp.189~200 (1997)
- Jyonouchi, H., "Imunomodulating actions of carotenoids: enhance-ment of in vivo and in vitro antibody production to T-dependent antigens", Nutr. Cancer, 21, pp.47~58 (1994)
- Lee, J.H, "Cloning and characteriza-tion of the astaxanthin biosynthesis gene cluster from the marine bacterium Paracoccus haeundaensis", Gene, (370), pp.86~95 (2006)
- Lee, J.H, "Functional expression of the astaxanthin biosynthesis genes from a marine bac-terium, Paracoccus haeundaensis", Biotechnol. Lett., 28, pp.1167~1173 (2006)
- Lee, J.H., "Paracoccus haeundaensis sp. nov., a Gram-negative, halophilic, astaxanthin-producing bacterium", Int. J. Syst. Evol. Microbiol., 54, pp.1699~1702 (2004)
- 김영태, "Functional Analysis of Combinations in Astaxanthin Biosynthesis Genes from Paracoccus haeundaensis", Biotechnology and Bioprocess Engineering, 12(3), pp.312~317 (2007), 한국생물공학회
- Math, S.K., "The crtE gene in Erwinia herbicola encodes geranylgeranyl diphosphate synthase", Proc. Natl. Acad. Sci. USA, 89, pp.6761~6764 (1992)
- Wang, K, "Chain-length determina-tion mechanism of isoprenyl diphosphate synthases and implications for molecular evolution", Trends Biochem. Sci., 24, pp.445~451 (1999)