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Effect of the oxygen transfer rate on oxygen-limited production of plasmid DNA by Escherichia coli

Lara, AR; Jaen, KE; Folarin, O; Keshavarz-Moore, E; Buechs, J; (2019) Effect of the oxygen transfer rate on oxygen-limited production of plasmid DNA by Escherichia coli. Biochemical Engineering Journal , 150 , Article 107303. 10.1016/j.bej.2019.107303. Green open access

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Abstract

The influence of oxygen limitation on pDNA yields in cultures of Escherichia coli was studied. Cultures at maximum oxygen transfer rate (OTR_{max}) values of 10, 14, 30 and 45 mmol L^{−1} h^{−1} and an aerobic culture at an OTR_{max} f 110 mmol L^{−1} h^{−1} were performed in microtiter plates (MTPs). Dissolved oxygen tension (DOT), pH, biomass and NADH fluorescence were monitored online. An E. coli strain that constitutively expresses Vitreoscilla hemoglobin (VHb) was used and compared with its parent strain. pDNA yields were inversely proportional to the OTR_{max} for both strains and increased more than two-fold in cultures at the lowest OTR_{max} compared with that in aerobic cultures. Expression of VHb increased the specific growth rate at OTR_{max} values of 10, 14 and 30 mmol L^{−1} h^{−1} compared with that of the parent strain. NADH-specific fluorescence decreased in cultures of the engineered strain. The pDNA supercoiled fraction (SCF) was greatest in cultures at an OTRmax of 30 mmol L^{−1} h^{−1{, compared with that of the parent strain. NADH-specific fluorescence decreased in cultures of the engineered strain. The pDNA supercoiled fraction (SCF) was greatest in cultures at an OTRmax of 30 mmol L−1 h−1, r

Type: Article
Title: Effect of the oxygen transfer rate on oxygen-limited production of plasmid DNA by Escherichia coli
Open access status: An open access version is available from UCL Discovery
DOI: 10.1016/j.bej.2019.107303
Publisher version: https://doi.org/10.1016/j.bej.2019.107303
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: Oxygen transfer rate, Plasmid DNA, Microbioreactors, Vitreoscilla hemoglobin, Scale-up/scale-down
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
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Chemical Engineering
URI: https://discovery.ucl.ac.uk/id/eprint/10110587
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