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Molecular cloning and characterization of farnesyl pyrophosphate synthase gene from Panax sokpayensis, a new Panax species from Sikkim Himalaya

B. Gurung,Savita Bains,D. Saha,Kashmir Singh,P. Bhardwaj,D. Sahoo

Published 2019 in Journal of Applied Research on Medicinal and Aromatic Plants

ABSTRACT

Abstract This study describes the molecular characterization and heterologous expression of farnesyl pyrophosphate synthase (PsFPS) from Panax sokpayensis Shiva K. Sharma & Pandit. FPS is a key regulatory enzyme of ginsenoside biosynthetic pathway that directs the metabolic flux from mevalonate (MVA) and methylerythritol phosphate (MEP) pathway towards the synthesis of ginsenosides and phytosterols. Using Rapid Amplification of cDNA Ends (RACE), full-length PsFPS of 1437 bp with an open reading frame (ORF) of 1026 bp coding for 342 amino acids was cloned. Deduced amino acid sequence of PsFPS contained two highly conserved aspartate rich motifs with consensus DDXXD sequence that constitute the main catalytic site of FPS. Full length cDNA of PsFPS has 5′ untranslated region (UTR) with 142 bp and 3′ UTR with 234 bp followed by a poly (A) tail. In silico analysis detected three identical polyadenylation signals (PAS) with canonical sequence, AATAAA in the 3′ UTR. Alignment of 3′ UTR of PsFPS with the FPS from other Panax species indicated the presence of alternative PAS which could have possible role in the formation of different isoforms of FPS in Panax species. Open reading frame of PsFPS was expressed in Escherichia coli (E. coli) and the recombinant protein was produced in vitro. High expression of recombinant protein was observed after 1 h of induction with 1 mM IPTG. HPLC mediated enzyme assay confirmed the production of functional PsFPS in E. coli. This is the first report on molecular characterization, heterologous expression and functional validation of PsFPS from P. sokpayensis.

PUBLICATION RECORD

  • Publication year

    2019

  • Venue

    Journal of Applied Research on Medicinal and Aromatic Plants

  • Publication date

    2019-09-01

  • Fields of study

    Biology, Chemistry, Environmental Science

  • Identifiers
    DOI 10.1016/J.JARMAP.2019.100215
  • External record

    Open on Semantic Scholar

  • Source metadata

    Semantic Scholar

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