Abstract
Plant cell suspension cultures of Thevetia peruviana has been explored for pharmaceutical compounds production. The aim of this study was to evaluate the agitation rate effect on growth and metabolism behavior of T. peruviana cells grown in a 7 L stirred tank reactor. Increases in agitation rates favored cell growth, secondary metabolites production and metabolic activity. The highest biomass concentration 11.92 ± 0.25 g DW/L was reached at 550 rpm. The oxidase-reductase activity was stimulated at 550 and 800 rpm. Guaiacol peroxidase activity showed an increase for 300 and 550 rpm after day 7. High levels of extracellular Reactive Oxygen Species (ROS) were observed at day 7 for 550 and 800 rpm. Intracellular phenolic compounds (PC) showed an upward trend until day 7 with a maximum phenolic production of 57.78 ± 4.70 mg EGA/100 g FW for 550 rpm. These results indicated that cells responded to ROS stress in a non-enzymatic manner during the first 7 days of culture, increasing PC production with antioxidant capacity. After 7 days, cells responded enzymatically. Intracellullar cardiac glycoside showed a relative increase of 1.7 and 2.1 times for 550 and 800 rpm, respectively. The maximum extracellular production of cardiac glycoside for 550 and 800 rpm was 770.34 ± 42.84 mg EP/L. Taken together this study established reactor culture conditions for production of cardiac glycosides and PC, especially taxifolin.
Key message
Cell suspensions of T. peruviana cultured in 7-liter reactor are tolerant at agitation rate up 800 rpm, maintaining biomass, cardiac glycosides, phenolic compounds and taxifolin production.
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Abbreviations
- Abs:
-
Absorbance units
- ABTS:
-
2,2-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)
- SH:
-
Schenk and Hildebrandt
- 2,4-D:
-
2,4-Dichlorophenoxyacetic acid
- DCF:
-
2,7-dichlorofluorescein
- DO:
-
Dissolved oxygen
- DW:
-
Dry weight
- EGA:
-
Equivalents gallic acid
- FDA:
-
Fluorescein diacetate
- FW:
-
Fresh weight
- GI:
-
Growth index
- GPX:
-
Guaiacol peroxidase activity
- HPLC–DAD:
-
High performance liquid chromatography—diode array detector
- MTT:
-
3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
- PC:
-
Phenolic compounds
- PVP:
-
Polyvinyl pyrrolidone
- RFU:
-
Relative fluorescence units
- RID:
-
Refractive index detector
- ROS:
-
Reactive oxigen species
- TEAC:
-
Trolox equivalent antioxidan capacity
- TPC:
-
Total phenols content
- YX/S :
-
Biomass substrate yield
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Acknowledgements
The authors would like to thanks to COLCIENCIAS - MINCIENCIAS for providing grants to Juan Pablo Arias and Dary Mendoza - National Program for Ph.D studies in Colombia, Announcements 567 and 673, respectively, and Postdoctoral Stays Program in entities of the National System of Science, Technology and Innovation (SNCTeI), Announcement 848 for Juan Pablo Arias – to the group of Química Orgánica de Productos Naturales of the Universidad de Antioquia for their collaboration in the identification of metabolites by mass spectroscopy.
Funding
This research was funded by Patrimonio Autónomo Fondo Nacional de Financiamiento para la Ciencia, la Tecnología y la Innovación Francisco José de Caldas - Departamento Administrativo de Ciencia, Tecnología e Innovación de Colombia – COLCIENCIAS, Grant Number FP44842-006-2018.
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JPA: Conceptualization, Methodology, Formal analysis, Investigation, Funding acquisition, Writing—Original Draft, Review & Editing, Project administration. DLM: Funding acquisition, Writing—Review & Editing. MA: Supervision, Funding acquisition, Writing—Review & Editing.
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Arias, J.P., Mendoza, D. & Arias, M. Agitation effect on growth and metabolic behavior of plant cell suspension cultures of Thevetia peruviana at bench scale reactor. Plant Cell Tiss Organ Cult 145, 307–319 (2021). https://doi.org/10.1007/s11240-021-02009-2
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DOI: https://doi.org/10.1007/s11240-021-02009-2