Production of recombinant butyrylcholinesterase from transgenic rice cell suspension cultures in a pilot‐scale bioreactor
| العنوان: | Production of recombinant butyrylcholinesterase from transgenic rice cell suspension cultures in a pilot‐scale bioreactor |
|---|---|
| المؤلفون: | Somen Nandi, Kantharakorn Macharoen, Karen A. McDonald, Min Du, Seongwon Jung |
| المصدر: | Biotechnology and Bioengineering. 118:1431-1443 |
| بيانات النشر: | Wiley, 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | chemistry.chemical_classification, Transgene, Oryza, Bioengineering, Plants, Genetically Modified, Plant cell, Applied Microbiology and Biotechnology, Genetically modified rice, Recombinant Proteins, law.invention, Enzyme, chemistry, Cell culture, law, Butyrylcholinesterase, Plant Cells, Bioreactor, Recombinant DNA, Food science, Sugar, Biotechnology |
| الوصف: | Producing recombinant proteins in transgenic plant cell suspension cultures in bioreactors provides controllability, reproducibility, scalability, and low-cost production, although low yields remain the major challenge. The studies on scaling-up to pilot-scale bioreactors, especially in conventional stainless-steel stirred tank bioreactors (STB), to produce recombinant proteins in plant cell suspension cultures are very limited. In this study, we scaled-up the production of rice recombinant butyrylcholinesterase (rrBChE), a complex hydrolase enzyme that can be used to prophylactically and therapeutically treat against organophosphorus nerve agents and pesticide exposure, from metabolically-regulated transgenic rice cell suspension cultures in a 40L pilot-scale STB. Employing cyclical operation together with a simplified-process operation (controlling gas sparging rate rather than dissolved oxygen and allowing natural sugar depletion) identified in lab-scale (5L) bioreactor studies, we found consistent maximum total active rrBChE production level of 46-58 µg/g fresh weight in four cycles over 82 days of continuous operation. Additionally, maintaining the overall volumetric oxygen mass transfer coefficient (kL a) in the pilot-scale STB to be equivalent to the lab-scale STB improves the maximum total active rrBChE production level and the maximum volumetric productivity to 85 µg/g fresh weight and 387 µg L-1 day-1 , respectively, which are comparable to the lab-scale culture. Here, we demonstrate pilot scale bioreactor performance using a metabolically-regulated transgenic rice cell culture for long-term, reproducible, and sustained production of rrBChE. This article is protected by copyright. All rights reserved. |
| تدمد: | 1097-0290 0006-3592 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::af52d96253a24e9f67c2ce7045430f44 https://doi.org/10.1002/bit.27638 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....af52d96253a24e9f67c2ce7045430f44 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10970290 00063592 |
|---|