title: Critical influence of Eschscholzia californica cells nutritional state on secondary metabolite production.
abstract: In this study, we investigated the influence of initial internal nutrient concentrations at the time of elicitation on the ability of Eschscholzia californica (EC) cells to produce alkaloids. Three EC cell suspensions cultivated in culture media differing in their PO4(3-) and NO3- contents were sampled daily for 12 days and analyzed for extracellular and intracellular nutrient concentrations. The ability of the cells to produce alkaloids was tested along the three cell suspension cultures. Sampled cells were then further cultured for 7 days in a production medium containing the elicitor and an extraction resin. The alkaloid production of the cells was measured 7 days post-elicitation. In the low-N medium, starch, glucose, and phosphate contents in the biomass was increased by 470, 1624 and 70%, respectively, 10 days after inoculation compared to the control culture in standard B5 medium. Cell concentration was significantly reduced from 10.3 to 8.6 millions cell/mL on this low-N medium compared to the control, nevertheless alkaloid production was multiplied by 39 at day 10 when cells were elicited. Cells grown on the low-N or low-P media accumulated 83% and 188% more carbon, respectively, than control cells at the end of the culture. This intracellular C was mainly stored in the form of starch in low-P medium and both in the form of starch and glucose in the low-N medium. The ability of EC cells to produce alkaloids upon elicitation was shown to be strongly dependent on the initial intracellular C and P content at the time of elicitation. This suggests that reproducibility and productivity during EC cell culture could be enhanced by manipulating the intracellular C and P content at the time of elicitation.
authors.affiliation: Canada Research Chair on the Development of Metabolic Engineering Tools Bio-P2 Research Unit, Department of Chemical Engineering, Ecole Polytechnique de Montréal, P.O. Box 6079, Centre-ville Station, Montréal, Québec, Canada H3C 3A7.
authors.forename: Laurence
authors.initials: L
authors.lastname: Lamboursain
authors.forename: Mario
authors.initials: M
authors.lastname: Jolicoeur
mesh_terms: D000470:Alkaloids / Q000096:biosynthesis*; D018533:Biomass; D002244:Carbon / Q000378:metabolism; D002452:Cell Count; D018929:Cell Culture Techniques; D002460:Cell Line; D003470:Culture Media / Q000737:chemistry*; D031675:Eschscholzia / Q000737:chemistry / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D009566:Nitrates; D010710:Phosphates / Q000032:analysis; D015203:Reproducibility of Results; D013213:Starch / Q000378:metabolism
chemical_list: D000470:Alkaloids; D003470:Culture Media; D009566:Nitrates; D010710:Phosphates; D002244:Carbon; D013213:Starch; D005947:Glucose