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A high-rate perfusion bioreactor for plant cells
C De Dobbeleer1, M Cloutier, M Fouilland
1Canada Research Chair on the Development of Metabolic Engineering Tools, Bio-P2, Department of Chemical Engineering, Ecole Polytechnique de Montreal, Station Centre-Ville, Montreal, Quebec, Canada H3C3A7.
Biotechnology and Bioengineering
|June 30, 2006
Summary
A novel perfusion bioreactor with continuous extraction capabilities was developed for Eschscholtzia californica cell cultures. While it maintained cell viability, secondary metabolite production was reduced compared to free resin methods, though chelilutine pathway was favored.
Area of Science:
- Biotechnology
- Plant Cell Culture
- Metabolic Engineering
Background:
- Secondary metabolites from plant cell cultures are valuable compounds.
- Efficient extraction methods are crucial for maximizing yields in bioreactor systems.
- Eschscholtzia californica produces valuable alkaloids, but optimizing their production remains a challenge.
Purpose of the Study:
- To design and evaluate a perfusion bioreactor system for continuous secondary metabolite extraction from Eschscholtzia californica cell suspensions.
- To assess the impact of continuous extraction on cell growth, viability, and alkaloid production.
- To compare the performance of the perfusion bioreactor with traditional culture methods.
Main Methods:
- A 2.5-L perfusion bioreactor equipped with sedimentation columns and XAD-7 resins was utilized.
- Eschscholtzia californica cells were cultured and elicited with chitin.
- Continuous extraction of medium from the bioreactor through fluidized resin columns was performed.
- Cellular growth rates and secondary metabolite production were quantified and compared to control cultures.
Main Results:
- The bioreactor maintained stable cell/medium separation with up to 90% sedimented cell volume (SCV).
- Specific growth rates remained consistent (0.24 +/- 0.04/day) before and after elicitation in perfusion cultures.
- Perfusion cultures showed reduced total alkaloid production (2.06 micromole/gDW) compared to free resin controls (30.94 micromole/gDW), attributed to nutritional state changes post-elicitation.
- The chelilutine production pathway was favored in the continuous extraction system.
Conclusions:
- The developed perfusion bioreactor enables continuous extraction of secondary metabolites from plant cell suspensions.
- Cell nutritional status post-elicitation significantly impacts secondary metabolite production in this system.
- Despite lower overall yields, the system shows potential for targeted metabolite production, favoring specific pathways like chelilutine.