Production of imidazole alkaloids in cell cultures of jaborandi as affected by the medium pH
The effect of pH (from 4.8 to 9.8) on the production of pilosine and pilocarpine and on their partition between cell and medium was studied in two lineages (P and PP) of Pilocarpus microphyllus cell suspension cultures. Highest mass accumulation was observed at high pHs and both lineages produced pilocarpine while only lineage PP produced pilosine. Both alkaloids were released in the medium but higher accumulation occurred in the cells. The highest production of pilocarpine was at pH 8.8–9.8 in both cell lineages. Other imidazole alkaloids were also identified in both lineages. At all pHs tested, the pH in the media cultures tended to stabilize around 6 after 10–15 days of cultivation. NO3 − and NH4 + variation in the media might partially explain the pH stabilization.
KeywordsMedium pH Membrane transporters Pilocarpine Pilocarpus microphyllus Pilosine
N.L.A., I.N.A. and A.C.H.F.S. thank the Conselho Nacional de Desenvolvimento Científico Tecnológico do Brasil (CNPq-Brasil) and the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) for student fellowships. M.N.E. and P.M. thank CNPq-Brasil for research fellowships. This work was partially supported by FAPESP.
- Abreu IN, Mazzafera P, Eberlin MN, Zullo MAT, Sawaya ACHF (2007b) Characterization of the variation of the imidazole alkaloid profile of Pilocarpus microphyllus in different seasons and parts of the plant by electrospray ionization mass spectrometry fingerprinting and identification of novel alkaloids by tandem mass spectrometry. Rapid Commun Mass Spectrom 21:1205–1213PubMedCrossRefGoogle Scholar
- Buckland E, Townsley PM (1975) Coffee cell suspension cultures: caffeine and chlorogenic acid content. J Inst Can Sci Technol 8:164–165Google Scholar
- Dong H, Zhong J (2001) Significant improvement of taxane cell production in suspension cultures of Taxus chinensis by combining elicitation with sucrose feed. Biotechnol Eng J 8:145–150Google Scholar
- Minocha SC (1987) pH of the medium and the growth and metabolism of cells in culture. In: Bonga JM, Durzan DJ (eds) Cell and tissue culture in forestry vol. 1: general principles and biotechnology. Martinus Nijhoff Publishers, Dordrecht, pp 125–141Google Scholar
- Pinheiro CUB (1997) Jaborandi (Pilocarpus sp., Rutaceae): a wild species and its transformation into a crop. Econ Bot 51:49–58Google Scholar
- Salisbury FB, Ross CW (1992) Plant physiology. Wadsworth Publishing Company, BelmontGoogle Scholar
- Wynn RL (1996) Oral pilocarpine (Salagen): a recently approved salivary stimulant. Gen Dent 44:29–30Google Scholar