Accumulation of amino acids deriving from pyruvate in Escherichia coli W3110 during fed-batch cultivation in a two-compartment scale-down bioreactor
Jaakko Soini, Kaisa Ukkonen, Peter Neubauer
DOI: 10.4236/abb.2011.25049   PDF    HTML     4,458 Downloads   8,859 Views   Citations


Bioprocess scale-down simulators are used to investigate the phenomena occurring in industrial scale cultivations. The aim is to simulate the large-scale conditions, characterized by long mixing times introducing local gradients into the bioreactor, in laboratory scale bioreactor. We have expanded the current physiological knowledge about the consequences of such cultivation conditions from well-known mixed-acid fermentation to amino acid metabolism. Several interesting deviations were observed for amino acids derived from pyruvate when a STR-PFR scale-down simulator and a regular STR fed-batch cultivation were compared. In this article we show that the cultivation of Escherichia coli K-12 substrain W3110 in a scale-down two-compartment bioreactor has consequences on the free amino acid pools. Repeated occurrence of high glucose zones affects the biosynthesis of amino acids derived from pyruvate. This is interesting since the pyruvate based amino acids are precursors of the branched chain amino acid pathway, which is further disturbed by an unbalanced regulation caused by a frameshift mutation in the ilvG gene of E. coli K-12 strains. As a consequence the formation of non-typical amino acid norvaline was detected in the two compartment bioreactor cultivation.

Share and Cite:

Soini, J. , Ukkonen, K. and Neubauer, P. (2011) Accumulation of amino acids deriving from pyruvate in Escherichia coli W3110 during fed-batch cultivation in a two-compartment scale-down bioreactor. Advances in Bioscience and Biotechnology, 2, 336-339. doi: 10.4236/abb.2011.25049.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Enfors, S.O., Jahic, M., Rozkov, A., Xu, B., Hecker, M., Jürgen, B., Krüger, E., Schweder, T., Hamer, G., O'Beirne, D., Noisommit-Rizzi, N., Reuss, M., Boone, L., Hewitt, C., McFarlane, C., Nienow, A., Kovacs, T., Tragardh, C., Fuchs, L., Revstedt, J., Friberg, P.C., Hjertager, B., Blomsten, G., Skogman, H., Hjort, S., Hoeks, F., Lin, H.Y., Neubauer, P., van der, L.R., Luyben, K., Vrabel, P. and Manelius, A. (2001) Physiological responses to mixing in large scale bioreactors. Journal of Biotechnology, 85,
[2] Lapin, A., Schmid, J. and Reuss, M. (2006) Modeling the dynamics of E-coli populations in the three-dimensional turbulent field of a stirred-tank bioreactor - A structured- segregated approach. Chemical Engineering Sci-ence, 61, 4783-4797.
[3] Chassagnole, C., Noisommit-Rizzi, N., Schmid, J.W., Mauch, K. and Reuss, M. (2002) Dynamic modeling of the central carbon metabolism of Escherichia coli. Biotechnology and Bioengineering, 79, 53-73.
[4] De Mey, M., Taymaz-Nikerel, H., Baart, G., Waegeman, H., Maertens, J., Heijnen, J.J. and van Gulik, W.M. (2010) Catching prompt metabolite dynamics in Escherichia coli with the BioScope at oxygen rich conditions. Metabolic Engineering, 12, 477-487.
[5] Lin, H.Y., Mathiszik, B., Xu, B., Enfors, S.O. and Neubauer, P. (2001) Determination of the maximum specific uptake capacities for glucose and oxygen in glucose- limited fed-batch cultivations of Escherichia coli. Biotechnology and Bioengineering, 73, 347-357.
[6] Neubauer, P., Haggstrom, L. and Enfors, S.O. (1995) Influence of substrate oscillations on acetate formation and growth yield in Escherichia coli glucose-limited fed-batch cultivations. Biotechnology and Bioengineering, 47, 139-146.
[7] Xu, B., Jahic, M., Blomsten, G. and Enfors, S.O. (1999) Glucose overflow metabolism and mixed-acid fermentation in aerobic large-scale fed-batch processes with Escherichia coli. Applied Microbiology and Biotechnology, 51, 564-571.
[8] Xu, B., Jahic, M. and Enfors, S.O. (1999) Modeling of overflow metabolism in batch and fed-batch cultures of Escherichia coli. Biotechnology Progress, 15, 81-90.
[9] Hewitt, C.J., Onyeaka, H., Lewis, G., Taylor, I.W. and Nienow, A.W. (2007) A comparison of high cell density fed-batch fermentations involving both induced and non-induced recombinant Escherichia coli under well-mixed small-scale and simulated poorly mixed large-scale conditions. Biotechnology and Bioengineering, 96, 495-505.
[10] Schaub, J., Mauch, K. and Reuss, M. (2008) Metabolic flux analysis in Escherichia coli by integrating isotopic dynamic and isotopic stationary 13C labeling data. Biotechnology and Bioengineering, 99, 1170-1185.
[11] Lara, A.R., Taymaz-Nikerel, H., Mashego, M.R., van Gulik, W.M., Heijnen, J.J., Ramirez, O.T. and van Winden, W.A. (2009) Fast dynamic response of the fermentative metabolism of Escherichia coli to aerobic and anaerobic glucose pulses. Biotechnology and Bioengineering, 104, 1153-1161.
[12] Soini, J., Falschlehner, C., Liedert, C., Bernhardt, J., Vuoristo, J. and Neubauer, P. (2008) Norvaline is accumulated after a down-shift of oxygen in Escherichia coli W3110. Microbial Cell Factories, 7, 30-
[13] Soini, J., Ukkonen, K. and Neubauer, P. (2008) High cell density media for Escherichia coli are generally designed for aerobic cultivations - consequences for large-scale bioprocesses and shake flask cultures. Microbial Cell Factories, 7, 26-
[14] George S, Larsson G and Enfors SO. (1993) A Scale-down 2-compartment reactor with controlled substrate oscillations - Metabolic response of Saccharomyces Cervisiae. Bioprocess Engineering, 9, 249-257.
[15] Neubauer, P., Ahman, M., Tornkvist, M., Larsson, G. and Enfors, S.O. (1995) Response of guanosine tetraphosphate to glucose fluctuations in fed-batch cultivations of Escherichia coli. Journal of Biotechnology, 43, 195-204.
[16] Andersen, D.C., Swartz, J., Ryll, T., Lin, N. and Snedecor, B. (2001) Metabolic oscillations in an E. coli fermentation. Biotechnology and Bioengineering, 75, 212-218.
[17] Apostol, I., Levine, J., Lippincott, J., Leach, J., Hess, E., Glascock, C.B., Weickert, M.J. and Blackmore, R. (1997) Incorporation of norvaline at leucine positions in recombinant human hemoglobin expressed in Escherichia coli. The Journal of Biological Chemistry, 272, 28980-28988.

Copyright © 2024 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.