Horticulture, Environment, and Biotechnology

, Volume 58, Issue 6, pp 613–619 | Cite as

Functional characterization of the β-amyrin synthase gene involved in platycoside biosynthesis in Platycodon grandiflorum

  • Yurry Um
  • Mei Lan Jin
  • Dae Young Lee
  • Chang Kug Kim
  • Chang Pyo Hong
  • Yi Lee
  • Ok Tae Kim
Research Report
  • 43 Downloads

Abstract

Platycodon grandiflorum A. DC. contains a variety of triterpene saponins induced from the base structure of oleanane-type in roots. The β-amyrin synthase (bAS) plays an important role in controlling the biosynthesis of triterpene saponins and we obtained the full-length cDNA of the gene, which is named PlgOSC1 in P. grandiflorum. To identify the PlgOSC1 products, we expressed it in heterologous yeast cells. Results of GC-MS analysis showed a unique peak that was consistent with that of the authentic β-amyrin standard with exactly the same retention time. Furthermore, the pattern of MS fragments of the peak was also exactly the same as that of the β-amyrin. In qRT-PCR analysis of four organs of P. grandiflorum, no difference in PlgOSC1 expression level was observed; therefore, platycosides derived from β-amyrin might be present in all P. grandiflorum tissues. In conclusion, PlgOSC1 encodes a bAS enzyme that catalyzes the first committed step of platycosides biosynthesis in P. grandiflorum.

Additional key words

beta-amyrin synthase oxidosqualene cyclase platycosides triterpenoids 

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Supplementary material

13580_2017_54_MOESM1_ESM.xlsx (14 kb)
Supplementary Table 1. Accession data for the OSCs used in the phylogenetic analyses.

References

  1. Abe I, Prestwich GD (1995) Identification of the active site of vertebrate oxidosqualene cyclase. Lipids 30:231–234CrossRefPubMedGoogle Scholar
  2. Andre CM, Legay S, Deleruelle A, Nieuwenhuizen N, Punter M, Brendolise C, Cooney JM, Lateur M, Hausman JF, et al (2016) Multifunctional oxidosqualene cyclases and cytochrome P450 involved in the biosynthesis of apple fruit triterpenic acids. New Phytol 211:1279–1294CrossRefPubMedPubMedCentralGoogle Scholar
  3. Dhar N, Rana S, Razdan S, Bhat WW, Hussain A, Dhar RS, Vaishnavi S, Hamid A, Vishwakarma R, et al (2014) Cloning and functional characterization of three branch point oxidosqualene cyclases from Withania somnifera (L.) dunal. J Biol Chem 289:17249–17267CrossRefPubMedPubMedCentralGoogle Scholar
  4. Ito R, Masukawa Y, Hoshino T (2013a) Purification, kinetics, inhibitors and CD for recombinant β-amyrin synthase from Euphorbia tirucalli L and functional analysis of the DCTA motif, which is highly conserved among oxidosqualene cyclases. FEBS J 280:1267–1280CrossRefPubMedGoogle Scholar
  5. Ito R, Hashimoto I, Masukawa Y, Hoshino T (2013b) Effect of cation-p interactions and steric bulk on the catalytic action of oxidosqualene cyclase: a case study of Phe728 of β-amyrin synthase from Euphorbia tirucalli L. Chemistry 19:17150–17158CrossRefPubMedGoogle Scholar
  6. Ito R, Masukawa Y, Nakada C, Amari K, Nakano C, Hoshino T (2014) β-Amyrin synthase from Euphorbia tirucalli. Steric bulk, not the p-electrons of Phe, at position 474 has a key role in affording the correct folding of the substrate to complete the normal polycyclization cascade. Org Biomol Chem 12:3836–3846PubMedGoogle Scholar
  7. Jin ML, Lee DY, Um Y, Lee JH, Park CG, Jetter R, Kim OT (2014) Isolation and characterization of an oxidosqualene cyclase gene encoding a β-amyrin synthase involved in Polygala tenuifolia Willd. saponin biosynthesis. Plant Cell Rep 33:511–519CrossRefPubMedGoogle Scholar
  8. Khan M, Maryam A, Zhang H, Mehmood T, Ma T (2016) Killing cancer with platycodin D through multiple mechanisms. J Cell Mol Med 20:389–402CrossRefPubMedGoogle Scholar
  9. Kushiro T, Shibuya M, Ebizuka Y (1998) β-amyrin synthase-Cloning of oxidosqualene cyclase that catalyzes the formation of the most popular triterpene among higher plants. Eur J Biochem 256:238–244CrossRefPubMedGoogle Scholar
  10. Lee JW, Ji SH, Kim GS, Song KS, Um Y, Kim OT, Lee Y, Hong CP, Shin DH, et al (2015) Global profiling of various metabolites in Platycodon grandiflorum by UPLC-QTOF/MS. Int J Mol Sci 16:26786–26796CrossRefPubMedPubMedCentralGoogle Scholar
  11. Liu Y, Zhao Z, Xue Z, Wang L, Cai Y, Wang P, Wei T, Gong J, Liu Z, et al (2016) An intronless β-amyrin synthase gene is more efficient in oleanolic acid accumulation than its paralog in Gentiana straminea. Sci Rep 6:33364CrossRefPubMedPubMedCentralGoogle Scholar
  12. Ma CH, Gao ZJ, Zhang JJ, Zhang W, Shao JH, Hai MR, Chen JW, Yang SC, Zhang GH (2016) Candidate genes involved in the biosynthesis of triterpenoid saponins in Platycodon grandiflorum identified by transcriptome analysis. Front Plant Sci 7:673PubMedPubMedCentralGoogle Scholar
  13. Misra RC, Maiti P, Chanotiya CS, Shanker K, Ghosh S (2014) Methyl jasmonate-elicited transcriptional responses and pentacyclic triterpene biosynthesis in sweet basil. Plant Physiol 164:1028–1044CrossRefPubMedGoogle Scholar
  14. Nyakudya E, Jeong JH, Lee NK, Jeong YS (2014) Platycosides from the roots of Platycodon grandiflorum and their health benefits. Prev Nutr Food Sci 19:59–68CrossRefPubMedPubMedCentralGoogle Scholar
  15. Phillips DR, Rasbery JM, Bartel B, Matsuda SP (2006) Biosynthetic diversity in plant triterpene cyclization. Curr Opin Plant Biol 9:305–314CrossRefPubMedGoogle Scholar
  16. Poralla K, Hewelt A, Prestwich GD, Abe I, Reipen I, Sprenger G (1994) A specific amino acid repeat in squalene and oxidosqualene cyclases. Trends Biochem Sci 19:157–158CrossRefPubMedGoogle Scholar
  17. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739CrossRefPubMedPubMedCentralGoogle Scholar
  18. Thimmappa R, Geisler K, Louveau T, O'Maille P, Osbourn A (2014) Triterpene biosynthesis in plants. Annu Rev Plant Biol 65:225–257CrossRefPubMedGoogle Scholar
  19. Wang Z, Yeats T, Han H, Jetter R (2010) Cloning and characterization of oxidosqualene cyclases from Kalanchoe daigremontiana: enzymes catalyzing up to 10 rearrangement steps yielding friedelin and other triterpenoids. J Biol Chem 285:29703–29712CrossRefPubMedPubMedCentralGoogle Scholar
  20. Wu Y, Zou HD, Cheng H, Zhao CY, Sun LF, Su SZ, Li SP, Yuan YP (2012) Cloning and characterization of a β-amyrin synthase gene from the medicinal tree Aralia elata (Araliaceae). Genet Mol Res 11:2301–2314CrossRefPubMedGoogle Scholar
  21. Zhang L, Wang Y, Yang D, Zhang C, Zhang N, Li M, Liu Y (2015) Platycodon grandiflorus - an ethnopharmacological, phytochemical and pharmacological review. J Ethnopharmacol 164:147–161CrossRefPubMedGoogle Scholar
  22. Zheng X, Luo X, Ye G, Chen Y, Ji X, Wen L, Xu Y, Xu H, Zhan R, et al (2015) Characterisation of two oxidosqualene cyclases responsible for triterpenoid biosynthesis in Ilex asprella. Int J Mol Sci 16:3564–3578CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Korean Society for Horticultural Science and Springer-Verlag GmbH Germany, part of Springer Nature 2017

Authors and Affiliations

  • Yurry Um
    • 1
  • Mei Lan Jin
    • 1
  • Dae Young Lee
    • 1
  • Chang Kug Kim
    • 2
  • Chang Pyo Hong
    • 3
  • Yi Lee
    • 4
  • Ok Tae Kim
    • 1
  1. 1.Department of Herbal Crop ResearchNational Institute of Horticultural and Herbal Science, RDAEumseongRepublic of Korea
  2. 2.Genomics DivisionNational Academy of Agricultural Science (NAAS), RDAJeonjuRepublic of Korea
  3. 3.Theragen Etex Bio Institute, Theragen EtexSuwonRepublic of Korea
  4. 4.Department of Industrial Plant Science and TechnologyChungbuk National UniversityCheongjuRepublic of Korea

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