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Fully integrated L-phenylalanine separation and concentration using reactive-extraction with liquid-liquid centrifuges in a fed-batch process with E. coli

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Abstract

A novel in situ product recovery (ISPR) approach for the (fully) integrated separation of L-phenylalanine (L-phe) from a 20 l fed-batch process with the recombinant L-tyrosine auxotrophic strain E. coli F-4/pF81 is presented. The strain was rationally constructed for the production of the aromatic amino acid. Glucose and tyrosine control is used. A reactive extraction system consisting of kerosene, the cation-selective carrier D2EHPA and sulphuric acid, all circulating in liquid-liquid centrifuges, is applied for the on-line L-phe separation from cell- and protein-free permeate. Permeate is drained off from the bioreactor bypass. Using the novel ISPR approach, a significantly extended product formation period at 0.25 mmol/(g*h) together with a reduced by-product formation and a 28% relative glucose/L-phe yield increase is observed. Thus, the ISPR approach is superior to the reference non-ISPR process and even offers extraction rates approximately three times higher than the published membrane-based process.

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Acknowledgements

The authors would like to thank Christine Kaldeweier, Andreas Franz and Heidi Haase-Reiff for their support during fermentation, data acquisition/process control and HPLC analysis. We are grateful to Ursula Degner and Johannes Bongaerts for their help with constructing strain F-4/pF81. The authors are also indebted to Prof. Christian Wandrey for offering optimum working conditions at the institute. They also wish to express their thanks to the staff of DSM/DSM Biotech GmbH for their fruitful cooperation and their financial co-support together with the German Federal Ministry of Education and Research (Grant no. 0311644).

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Authors and Affiliations

  1. Institute of Biotechnology, Forschungszentrum Jülich GmbH, 52425, Jülich, Germany

    N. Rüffer, U. Heidersdorf, I. Kretzers, G. A. Sprenger & R. Takors

  2. Institute of Microbiology, University of Stuttgart, Allmandring 31, 70569, Stuttgart, Germany

    G. A. Sprenger

  3. DSM Biotech GmbH, Karl-Heinz Beckurts Str. 13, 52428, Jülich, Germany

    L. Raeven

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  1. N. Rüffer
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  2. U. Heidersdorf
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  3. I. Kretzers
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  4. G. A. Sprenger
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  5. L. Raeven
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  6. R. Takors
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Correspondence to R. Takors.

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Rüffer, N., Heidersdorf, U., Kretzers, I. et al. Fully integrated L-phenylalanine separation and concentration using reactive-extraction with liquid-liquid centrifuges in a fed-batch process with E. coli . Bioprocess Biosyst Eng 26, 239–248 (2004). https://doi.org/10.1007/s00449-004-0354-4

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  • DOI: https://doi.org/10.1007/s00449-004-0354-4

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