Cloning and characterization of the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway genes of a natural-rubber producing plant, Hevea brasiliensis

Tomoki Sando, Shinya Takeno, Norie Watanabe, Hiroshi Okumoto, Tomohisa Kuzuyama, Atsushi Yamashita, Masahira Hattori, Naotake Ogasawara, Eiichiro Fukusaki, Akio Kobayashi

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Abstract

Natural rubber is synthesized as rubber particles in the latex, the fluid cytoplasm of laticifers, of Hevea brasiliensis. Although it has been found that natural rubber is biosynthesized through the mevalonate pathway, the involvement of an alternative 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway is uncertain. We obtained all series of the MEP pathway candidate genes by analyzing expressed sequence tag (EST) information and degenerate PCR in H. brasiliensis. Complementation experiments with Escherichia coli mutants were performed to confirm the functions of the MEP pathway gene products of H. brasiliensis together with those of Arabidopsis thaliana, and it was found that 1-deoxy-D-xylulose-5-phosphate reductoisomerase, 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase, and 2-C-methyl-D-erythritol 2,4-cyclodiphosphate synthase of H. brasiliensis were functionally active in the E. coli mutants. Gene expression analysis revealed that the expression level of the HbDXS2 gene in latex was relatively high as compared to those of other MEP pathway genes. However, a feeding experiment with [1-13C] 1-deoxy-D-xylulose triacetate, an intermediate derivative of the MEP pathway, indicated that the MEP pathway is not involved in rubber biosynthesis, but is involved in carotenoids biosynthesis in H. brasiliensis.

Original languageEnglish
Pages (from-to)2903-2917
Number of pages15
JournalBioscience, Biotechnology and Biochemistry
Volume72
Issue number11
DOIs
Publication statusPublished - 2008
Externally publishedYes

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Keywords

  • 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway
  • cis-polyisoprene
  • Hevea brasiliensis
  • Natural rubber
  • Rubber biosynthesis

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Applied Microbiology and Biotechnology
  • Analytical Chemistry
  • Organic Chemistry

Cite this

Sando, T., Takeno, S., Watanabe, N., Okumoto, H., Kuzuyama, T., Yamashita, A., Hattori, M., Ogasawara, N., Fukusaki, E., & Kobayashi, A. (2008). Cloning and characterization of the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway genes of a natural-rubber producing plant, Hevea brasiliensis. Bioscience, Biotechnology and Biochemistry, 72(11), 2903-2917. https://doi.org/10.1271/bbb.80387