Biosynthesis stimulation of nor-secotriterpene anxiolytics in cell suspension cultures of Galphimia glauca Cav

Authors

  • Lidia Osuna,

    Corresponding author
    1. Centro de Investigación Biomédica del Sur, Instituto Mexicano del Seguro Social (CIBIS-IMSS), Xochitepec, Morelos, México
    • Correspondence: Prof. Lidia Teresa Osuna Torres (osunalidia@yahoo.com), Centro de Investigación Biomédica del Sur, Instituto Mexicano del Seguro Social, Argentina 1, 62790 Col. Centro, Xochitepec, Morelos, México

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  • Nadia Tapia,

    1. Centro de Investigación Biomédica del Sur, Instituto Mexicano del Seguro Social (CIBIS-IMSS), Xochitepec, Morelos, México
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  • Alejandro Zamilpa,

    1. Centro de Investigación Biomédica del Sur, Instituto Mexicano del Seguro Social (CIBIS-IMSS), Xochitepec, Morelos, México
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  • Enrique Jiménez-Ferrer,

    1. Centro de Investigación Biomédica del Sur, Instituto Mexicano del Seguro Social (CIBIS-IMSS), Xochitepec, Morelos, México
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  • Jaime Tortoriello

    1. Centro de Investigación Biomédica del Sur, Instituto Mexicano del Seguro Social (CIBIS-IMSS), Xochitepec, Morelos, México
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Abstract

Galphimia glauca produces compounds denominated galphimines (galphimine-A, galphimine-B and galphimine-E). Due to their important anxiolytic activity, we initiated in vitro cultures of the species with the purpose of developing a biotechnological process for obtaining galphimines. In this work, we stimulated the biosynthesis and excretion of galphimines with two-phase batch-type cell suspension cultures of G. glauca. The effect of nutritional variation and the 2,4-dichlorophenoxy acetic acid added to Murashige & Skoog(MS) culture medium was evaluated. Later, we evaluated the effect of the stimulation with calcium and methyl jasmonate (MeJ). The greatest production of galphimine-B (3.39 × 10−5 g/L day−1) was obtained on day 40 of kinetics, and induced by a treatment containing concentrations of nitrates and phosphate that are double of those normally used in MS medium, without sucrose but with added 2,4-dichlorophenoxy acetic acid (4 mg/L). Time of galphimine-B biosynthesis diminished due to the effect of MeJ in combination with calcium, and induced the excretion (100%) of galphimine-B (6.35 × 10−5 g/L day−1) into the culture medium. Thus, the use of calcium and MeJ comprises a viable alternative to stimulate the production and excretion of galphimine-B and galphimine-A in batch-type cultures of G. glauca in modified MS medium. Once optimized, the production of the anxiolytic compounds can be scaled up to the industrial level.

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