Microbial nar-GFP cell sensors reveal oxygen limitations in highly agitated and aerated laboratory-scale fermentors |
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| Authors: | Jose R Garcia Hyung J Cha Govind Rao Mark R Marten William E Bentley |
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| Institution: | (1) Fischell Department of Bioengineering, University of Maryland, 20742 College Park, MD, USA;(2) Department of Chemical and Biomolecular Engineering, University of Maryland, 20742 College Park, MD, USA;(3) Department of Chemical Engineering, Pohang University of Science and Technology, 790-784 Pohang, Korea;(4) Department of Chemical and Biochemical Engineering, University of Maryland Baltimore County, 21052 Baltimore, MD, USA |
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| Abstract: | Background Small-scale microbial fermentations are often assumed to be homogeneous, and oxygen limitation due to inadequate micromixing
is often overlooked as a potential problem. To assess the relative degree of micromixing, and hence propensity for oxygen
limitation, a new cellular oxygen sensor has been developed. The oxygen responsive E. coli nitrate reductase (nar) promoter was used to construct an oxygen reporter plasmid (pNar-GFPuv) which allows cell-based reporting of oxygen limitation.
Because there are greater than 109 cells in a fermentor, one can outfit a vessel with more than 109 sensors. Our concept was tested in high density, lab-scale (5 L), fed-batch, E. coli fermentations operated with varied mixing efficiency – one verses four impellers. |
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