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Control of endotoxin release in Escherichia coli fed-batch cultures

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Abstract

High amounts of outer membrane (OM) components were released in glucose-limited fed-batch (GLFB) cultures at 37 °C at specific growth rates approaching 0.05 h−1. Endotoxin analyses from a 20 °C GLFB culture gave similar results. An alternative fermentation technique, the temperature-limited fed-batch (TLFB) technique, reduced the endotoxin concentration in a culture with a biomass concentration of 30 g l−1 from the 850 mg l−1 in traditional GLFB cultures to about 20 mg l−1. The TLFB technique uses the temperature to regulate the dissolved oxygen tension, while all substrate components are unregulated. It appears to be severe glucose limitation that triggers the extensive release of endotoxins rather than a low growth rate. Furthermore, it is not the low temperature that stabilizes the OM when using the TLFB technique. Simulations and experimental data show that this technique results in the same biomass productivity as the GLFB technique.

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Abbreviations

LPS:

Lipopolysaccharide

OM:

Outer membrane

GLFB:

Glucose limited fed-batch

TLFB:

Temperature limited fed-batch

EU:

Endotoxin unit

DOT:

Dissolved oxygen tension

OD:

Optical density

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Acknowledgement

This work was supported by a grant from the Swedish Centre for Bioprocess Tecnology, CBioPT.

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Authors and Affiliations

  1. Department of Biotechnology, Royal Institute of Technology (KTH), Roslagstullsbacken 21, 10691, Stockholm, Sweden

    Marie Svensson, Ling Han, Gustav Silfversparre, Lena Häggström & Sven-Olof Enfors

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  1. Marie Svensson
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  2. Ling Han
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  3. Gustav Silfversparre
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  4. Lena Häggström
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  5. Sven-Olof Enfors
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Correspondence to Sven-Olof Enfors.

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Svensson, M., Han, L., Silfversparre, G. et al. Control of endotoxin release in Escherichia coli fed-batch cultures. Bioprocess Biosyst Eng 27, 91–97 (2005). https://doi.org/10.1007/s00449-004-0377-x

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  • DOI: https://doi.org/10.1007/s00449-004-0377-x

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