Thursday, 2 June 2016

Hyperforin production in Hypericum perforatum root cultures

Journal of Biotechnology
Volume 222, 20 March 2016, Pages 47–55

Mariam Gaida, c, ,

doi:10.1016/j.jbiotec.2016.02.016
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Highlights

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Root cultures of the medicinal plant Hypericum perforatum produce hyperforins, unlike roots of field-grown plants.
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The cultivated roots lack the photosensitizing hypericins.
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The root cultures provide an alternative production platform for hyperforins.

Abstract

Extracts of the medicinal plant Hypericum perforatum are used to treat depression and skin irritation. A major API is hyperforin, characterized by sensitivity to light, oxygen and temperature. Total synthesis of hyperforin is challenging and its content in field-grown plants is variable. We have established in vitro cultures of auxin-induced roots, which are capable of producing hyperforin, as indicated by HPLC-DAD and ESI–MS analyses. The extraction yield and the productivity upon use of petroleum ether after solvent screening were ∼5 mg/g DW and ∼50 mg/L culture after six weeks of cultivation. The root cultures also contained secohyperforin and lupulones, which were not yet detected in intact plants. In contrast, they lacked another class of typical H. perforatum constituents, hypericins, as indicated by the analysis of methanolic extracts. Hyperforins and lupulones were stabilized and enriched as dicyclohexylammonium salts. Upon up-scaling of biomass production and downstream processing, H. perforatum root cultures may provide an alternative platform for the preparation of medicinal extracts and the isolation of APIs.

Abbreviations

  • API, active pharmaceutical ingredient;
  • AUC, area under the curve;
  • DCHA, dicyclohexylamine;
  • EPI, enhanced product ion scan;
  • ESI, electrospray ionization;
  • HPLC-DAD, high performance liquid-chromatography-diode array detection;
  • IAA, indole-3-acetic acid;
  • IBA, indole-3-butyric acid;
  • ESI–MS, electrospray ionization–mass spectrometry;
  • ESI–MS/MS, electrospray ionization–tandem mass spectrometry;
  • MS, Murashige-Skoog medium;
  • PVDF, polyvinylidenfluoride;
  • RP, reverse phase;
  • TRPC6, transient receptor potential channel 6;
  • Ye, extraction yield

Chemical compounds studied in this article

  • Hyperforin (PubChem CID: 441298);
  • Hyperforin dicyclohexylammonium salt (PubChem CID: 46926346);
  • Adhyperforin (PubChem CID: 90658004);
  • Secohyperforin (PubChem CID: 101442297);
  • Adsecohyperforin (PubChem CID: 101442298);
  • Lupulone (PubChem CID: 68051);
  • Adlupulone (PubChem CID: 9909740)

Keywords

  • Hyperforins;
  • Hypericins;
  • Hypericum perforatum;
  • Lupulones;
  • Root cultures
Corresponding author at: Mendelssohnstr 1, 38106 Braunschweig, Germany.
Copyright © 2016 Elsevier B.V. All rights reserved.
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