loading page

Production of Recombinant Butyrylcholinesterase from Transgenic Rice Cell Suspension Cultures in a pilot scale bioreactor
  • +2
  • Kantharakorn Macharoen,
  • Min Du,
  • Seongwon Jung,
  • Karen McDonald,
  • Somen Nandi
Kantharakorn Macharoen
University of California Davis

Corresponding Author:[email protected]

Author Profile
Min Du
University of California Davis
Author Profile
Seongwon Jung
University of California Davis
Author Profile
Karen McDonald
University of California, Davis, University of California, Davis
Author Profile
Somen Nandi
University of California, Davis
Author Profile

Abstract

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 40-L 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 (5-L) 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 (kLa) 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.
25 Aug 2020Submitted to Biotechnology and Bioengineering
25 Aug 2020Submission Checks Completed
25 Aug 2020Assigned to Editor
01 Sep 2020Reviewer(s) Assigned
11 Oct 2020Review(s) Completed, Editorial Evaluation Pending
11 Oct 2020Editorial Decision: Revise Major
13 Nov 20201st Revision Received
13 Nov 2020Submission Checks Completed
13 Nov 2020Assigned to Editor
17 Nov 2020Review(s) Completed, Editorial Evaluation Pending
17 Nov 2020Editorial Decision: Accept
Apr 2021Published in Biotechnology and Bioengineering volume 118 issue 4 on pages 1431-1443. 10.1002/bit.27638