Design and Characterization of an Osmotic Pump System for Optimal Feeding and pH Control in E. coli Culture to Increase Biomass.
| Autor: | Abedin S; Department of Pharmaceutics and Pharmaceutical Nanotechnology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran., Ranjbari J; Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran., Haeri A; Department of Pharmaceutics and Pharmaceutical Nanotechnology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran., Vahidi H; Department of Pharmaceutics and Pharmaceutical Nanotechnology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran., Moghimi HR; Department of Pharmaceutics and Pharmaceutical Nanotechnology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran. |
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| Jazyk: | angličtina |
| Zdroj: | Iranian journal of pharmaceutical research : IJPR [Iran J Pharm Res] 2024 Feb 17; Vol. 23 (1), pp. e138677. Date of Electronic Publication: 2024 Feb 17 (Print Publication: 2024). |
| DOI: | 10.5812/ijpr-138677 |
| Abstrakt: | Background: Batch cultures used for various purposes, such as expression screening and recombinant protein production in laboratories, usually have some drawbacks due to the bolus addition of carbon sources, such as glucose and buffers, that lead to overflow metabolism, decreased pH, high osmolality, low biomass yield, and low protein production. Objectives: This study aimed to overcome the problems of batch culture using the controlled release concept by a controlled porosity osmotic pump (CPOP) system. Methods: The CPOP was formulated with glucose as a carbon source feeding and sodium carbonate as a pH modifier in the core of the tablet that was coated with a semipermeable membrane containing cellulose acetate and polyethylene glycol (PEG) 400. The release rate was regulated with Eudragit L100 as a retardant agent in the core and PEG 400 as a pore-former agent in the coating membrane. Fourier-transform infrared spectroscopy (FT-IR) and scanning electron microscopy (SEM) were used to elucidate compatibility between components and release mechanism, respectively. The in-vitro release of glucose and Na Results: Fourier-transform infrared spectroscopy confirmed the compatibility between the components in the osmotic pump, and SEM elucidated the release mechanism due to in-situ delivery pores created by dissolving soluble components (PEG 400) on the coated membrane upon contact with the dissolution medium. The in-vitro release studies indicated that the osmotic pump was able to deliver glucose and sodium carbonate in a zero-order manner. The use of CPOP in E. coli (BL21) cultivation resulted in a statistically significant improvement in biomass (over 80%), maintaining the pH of the medium (above 6.8) during the exponential phase, and reducing metabolic by-product formation (acetate), compared to bolus feeding (P < 0.05). Conclusions: The use of CPOP, which is capable of controlled release of glucose as a carbon source and sodium carbonate as a pH modifier, can overcome the drawbacks of bolus feeding, such as decreased pH, increased acetate concentration, and low productivity. It has a good potential for commercialization. Competing Interests: This work is a part of Saeedeh Abedin’s PhD thesis at the School of Pharmacy, Shahid Beheshti University of Medical Sciences (SBMU), Tehran, Iran. All the authors are academic members of SBMU, and their roles in the thesis are either as supervisors (HM, JR, and HV) or consulting supervisors (AH). This work was financially supported only by a grant from SBMU. Among the authors, HV and HM are members of the editorial board of this journal, and AH is one of the reviewers of the journal. There is no personal financial interest or any company related to this work. No patent has been published for this work yet. No fee is paid to anyone for consultation regarding this work. (Copyright © 2024, Abedin et al.) |
| Databáze: | MEDLINE |
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