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Construction of an l-phenylalanine-producing tyrosine-prototrophic Escherichia coli strain using tyrA ssrA-like tagged alleles

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Abstract

To construct a Phe-producing Tyr+ Escherichia coli strain, TyrA (chorismate mutase/prephenate dehydrogenase) activity was varied by engineering a proteolytically unstable protein. The tyrA in the E. coli BW25113 was altered to include ssrA-like tags. The tagged tyrA genes, which ensured different growth rates in M9 medium, were introduced into a Phe-producing strain to replace ΔtyrA. Strains with unstable TyrA-(A)ANDENYALAA proteins had a lower biomass yield and a higher Phe accumulation than strains generating the more stable TyrA-(A)ANDENYALDD. The Tyr/Phe ratio produced by the TyrA-tag strains was 10-fold less than that produced by the TyrAwt strain.

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Authors and Affiliations

  1. Ajinomoto-Genetika Research Institute, Moscow, 117545, Russian Federation

    Vera G. Doroshenko, Svetlana M. Kazakova, Alexander D. Kivero, Tatyana A. Yampolskaya & Sergey V. Mashko

  2. State Research Institute of Genetics and Selection of Industrial Microorganisms, Moscow, 117545, Russian Federation

    Rustem S. Shakulov

Authors
  1. Vera G. Doroshenko
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  2. Rustem S. Shakulov
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  3. Svetlana M. Kazakova
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  4. Alexander D. Kivero
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  5. Tatyana A. Yampolskaya
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  6. Sergey V. Mashko
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Corresponding author

Correspondence to Vera G. Doroshenko.

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Doroshenko, V.G., Shakulov, R.S., Kazakova, S.M. et al. Construction of an l-phenylalanine-producing tyrosine-prototrophic Escherichia coli strain using tyrA ssrA-like tagged alleles. Biotechnol Lett 32, 1117–1121 (2010). https://doi.org/10.1007/s10529-010-0265-1

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  • DOI: https://doi.org/10.1007/s10529-010-0265-1

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