Wang, B;
Nesbeth, D;
Keshavarz-Moore, E;
(2019)
Sorbitol/methanol mixed induction reduces process impurities and improves centrifugal dewatering in Pichia pastoris culture.
Enzyme and Microbial Technology
, 130
, Article 109366. 10.1016/j.enzmictec.2019.109366.
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Keshavarz-Moore_Sorbitol:methanol mixed induction reduces process impurities and improves centrifugal dewatering in Pichia pastoris culture_AAM.pdf - Accepted Version Download (1MB) | Preview |
Abstract
This study investigates how sorbitol/methanol mixed induction affects fermentation performance, dewatering characteristics of cells during harvesting and the profile of host cell proteins (HCP) in the process fluid when producing the target recombinant protein aprotinin. Compared to standard methanol induction, sorbitol/methanol (1:1, C-mol/C-mol) mixed induction improved cellular viability from 92.8 ± 0.3% to 97.7 ± 0.1% although resulted in a reduced product yield from 1.65 ± 0.03 g L^{-1} 1.12 ± 0.07 g L^{-1}. On the other hand, average oxygen consumption rate (OUR) dropped from 241.4 ± 21.3 mmol L^{-1} h^{-1} to 145.5 ± 6.7 mmol L^{-1} h^{-1}. Cell diameter decreased over time in the mixed induction, resulting in a D_{50} value of 3.14 μm at harvest compared to 3.85 μm with methanol. The reduction in cell size enhanced the maximum dewatering efficiency from 78.1 ± 3.9% to 84.5 ± 3.3% as evaluated by using an established ultra scale-down methodology that models pilot and industrial scale disc stack centrifugation. Seventy host cell proteins (HCPs) were identified in clarified supernatant when using sorbitol/methanol mixed induction regimen. The total number of HCPs identified with standard methanol induction was nearly one hundred. The downstream process advantage of the mixed induction lies in improved product purity by reducing both cell mortality and level of released whole cell proteins. This needs to be balanced and optimised against the observed reduction in product yield during fermentation.
| Type: | Article |
|---|---|
| Title: | Sorbitol/methanol mixed induction reduces process impurities and improves centrifugal dewatering in Pichia pastoris culture |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1016/j.enzmictec.2019.109366 |
| Publisher version: | https://doi.org/10.1016/j.enzmictec.2019.109366 |
| Language: | English |
| Additional information: | This version is the author accepted manuscript. For information on re-use, please refer to the publisher's terms and conditions. |
| Keywords: | P. pastoris fermentation, Sorbitol/methanol mixed induction, Dewatering, Host cell proteins |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > UCL BEAMS UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10110586 |
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