Biosynthesis stimulation of nor‐secotriterpene anxiolytics in cell suspension cultures of G alphimia glauca C av

G alphimia 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 M urashige & S koog (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 M e J 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 M e J 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.