IPTG limitation avoids metabolic burden and acetic acid accumulation in induced fed-batch cultures of Escherichia coli M15 under glucose limiting conditions |
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| Institution: | 1. Department of Urology, University Hospital Basel, Basel, Switzerland;2. Laboratory of Biomechanics and Biocalorimetry (LOB2), University of Basel, Basel, Switzerland;1. Christian Doppler Laboratory for Mechanistic and Physiological Methods for Improved Bioprocesses, Institute of Chemical Environmental and Bioscience Engineering, Vienna University of Technology, Vienna, Austria;2. Research Division Biochemical Engineering, Institute of Chemical Environmental and Bioscience Engineering, Vienna University of Technology, Vienna, Austria;1. Department of Biotechnology and Environmental Chemistry, Kitami Institute of Technology, 165 Koen-cho, Kitami, Hokkaido 090-8507, Japan;2. Department of Biotechnology and Environmental Chemistry, Graduate School of Engineering, Kitami Institute of Technology, 165 Koen-cho, Kitami, Hokkaido 090-8507, Japan;3. Biotechnology and Food Chemistry Course Program, School of Regional Innovation and Social Design Engineering, Kitami Institute of Technology, 165 Koen-cho, Kitami, Hokkaido 090-8507, Japan;1. Institute of Chemical, Environmental and Biological Engineering, TU Wien, Gumpendorfer Straße 1a, 1060 Wien, Austria;2. CD Laboratory on Mechanistic and Physiological Methods for Improved Bioprocesses, TU Wien, Gumpendorfer Straße 1a, 1060 Wien, Austria;1. Protein Research Chair, Department of Biochemistry, College of Science, King Saud University, PO Box 2455, Riyadh 11451, Saudi Arabia;2. Genome Research Chair, Department of Biochemistry, College of Science, King Saud University, PO Box 2455, Riyadh 11451, Saudi Arabia;3. Department of Basic Sciences, College of Science and Health Professions, King Saud Bin Abdulaziz University for Health Sciences, PO Box 3660, Riyadh 11426, Saudi Arabia;4. Biochemistry Department, Faculty of Science, Ain-Shams University, Abbassia 11381, Cairo, Egypt |
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| Abstract: | The most common strategy to produce recombinant proteins using Escherichia coli as expression vector is fed-batch culture, since high cell density cultures strategies have successfully been applied. Several methodologies to limit the specific growth rate in order to control E. coli metabolism have been defined, demonstrating that cultures can be grown under glucose limitation up to high cell densities without accumulation of acetic acid. However, under induction conditions it has been observed that E. coli metabolism is reorganized again and leads to acetic acid accumulation, causing inhibition of cell growth and decreasing protein expression efficiency.We propose a double limitation strategy (glucose and IPTG) for E. coli fed-batch cultures to avoid the deregulation of the metabolism in the induction phase. Reducing the concentration of IPTG while keeping glucose growth limitation, the accumulation of acetic acid decreased. At an IPTG concentration of 0.03 mmol/g DCW no accumulation of acetic acid was observed during the induction phase, in contraposition to what has normally been observed.Although a slight reduction of protein expression rate was observed when applying this double limitation strategy, the bioprocess volumetric productivity was enhanced due to the capability to prolong the induction phase, reaching higher levels of protein production. Another advantage of this strategy is the reduction of media cost due to the lower level of IPTG used. |
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