Molecular cloning, expression and characterization of cDNa encoding cis-prenyltransferases from Hevea brasiliensis: A key factor participating in natural rubber biosynthesis

Kasem Asawatreratanakul, Yuan Wei Zhang, Dhirayos Wititsuwannakul, Rapepun Wititsuwannakul, Seiji Takahashi, Atiya Rattanapittayaporn, Tanetoshi Koyama

Research output: Contribution to journalArticlepeer-review

121 Citations (Scopus)

Abstract

Natural rubber from Hevea brasiliensis is a high molecular mass polymer of isoprene units with cis-configuration. The enzyme responsible for the cis-1,4-polymerization of isoprene units has been identified as a particle-bound rubber transferase, but no gene encoding this enzyme has been cloned from rubber-producing plants. By using sequence information from the conserved regions of cis-prenyl chain elongating enzymes that were cloned recently, we have isolated and characterized cDNAs from H. brasiliensis for a functional factor participating in natural rubber biosynthesis. Sequence analysis revealed that all of the five highly conserved regions among cis-prenyl chain elongating enzymes were found in the protein sequences of the Hevea cis-prenyltransferase. Northern blot analysis indicated that the transcript(s) of the Hevea cis-prenyltransferase were expressed predominantly in the latex as compared with other Hevea tissues examined. In vitro rubber transferase assays using the recombinant gene product overexpressed in Escherichia coli revealed that the enzyme catalyzed the formation of long chain polyprenyl products with approximate sizes of 2 × 103-1 × 104 Da. Moreover, in the presence of washed bottom fraction particles from latex, the rubber transferase activity producing rubber product of high molecular size was increased. These results suggest that the Hevea cis-prenyltransferase might require certain activation factors in the washed bottom fraction particles for the production of high molecular mass rubber.

Original languageEnglish
Pages (from-to)4671-4680
Number of pages10
JournalEuropean Journal of Biochemistry
Volume270
Issue number23
DOIs
Publication statusPublished - 2003 Dec

Keywords

  • Hevea brasiliensis
  • Isoprenoid
  • Prenyltransferase
  • Rubber transferase

ASJC Scopus subject areas

  • Biochemistry

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