Shear rate as scale-up criterion of the protein production with enhanced proteolytic activity by phosphate addition in the Jacaratia mexicana cell culture

Abstract

The main challenge in bioreactor scale-up is maintaining the bioprocess productivity. The aim of the present work was to scale-up the protein production with enhanced proteolytic activity by phosphate addition in Jacaratia mexicana cell culture, by using the shear rate as scale-up criterion. The volumetric proteolytic activity in the J. mexicana cell culture in shake flask increased 1.9 times with increasing phosphate concentration from 1.25 to 2.5 mmol L⁻¹, while the protein concentration was similar (∼55 mg L⁻¹). This culture was scaled-up from 0.4-L to 4-L in stirred tank bioreactors at average shear rate of 200 s⁻¹ and 234 s⁻¹, respectively, increasing the phosphate concentration in the 4-L bioreactor (2.5 mmol L⁻¹) with respect to the 0.4-L bioreactor (1.25 mmol L⁻¹). There was also an increase in the volumetric proteolytic activity (2 times), without statistically significant difference in the protein concentration. Scale-up strategy and improvement of the culture medium exhibited a similar behaviour maintaining the protein productivity and increasing the proteolytic activity. These results indicate that the average shear rate is a useful hydrodynamic parameter in the scale-up of protein production of J. mexicana cell culture. Additional phosphate and bioreactor hydrodynamics could be related to unusual lengthening and thinning of most cells in the 4-L stirred tank bioreactor. Finally, the power input per unit volume in the scaled-up bioreactor decreased by 53%, improving the volumetric proteolytic activity and maintaining protein productivity. These results encourage proceeding with the scale-up process using the shear rate as a scale-up criterion.

Keywords:

• Stirred tank bioreactor
• protein production
• hydrodynamic parameters
• average shear rate
• proteolytic activity
• plant cell culture

Acknowledgements

The authors wish to acknowledge the financial support by Instituto Politécnico Nacional (IPN), México SIP 20195364 and 20200740. C. Oliver-Salvador and S. García-Salas are recipients of a fellowship from Comisión de Operación y Fomento de Actividades Académicas-IPN. Gómez-Montes E. O. was a recipient of a posdoctoral fellowship from the Estancias Posdoctorales Nacionales del Consejo Nacional de Ciencia y Tecnología CONACYT, México.

Disclosure statement

The authors report no conflict of interest

Data availability statement

All data that support the findings reported in this study are available from the corresponding author upon reasonable request.

Author contributions

García-Salas: Conceptualization, Data analysis and interpretation, Writing – review & editing. Gómez-Montes: Experiment conducting, data collect, statistical analysis, drafting the manuscript. Ramírez-Sotelo: Data analysis, Drafting the manuscript and review the manuscript. Oliver-Salvador: Concept and design, Data analysis and interpretation, drafting the manuscript & review