Biotechnology Letters

, Volume 32, Issue 8, pp 1117–1121

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

  • Vera G. Doroshenko
  • Rustem S. Shakulov
  • Svetlana M. Kazakova
  • Alexander D. Kivero
  • Tatyana A. Yampolskaya
  • Sergey V. Mashko
Original Research Paper

DOI: 10.1007/s10529-010-0265-1

Cite this article as:
Doroshenko, V.G., Shakulov, R.S., Kazakova, S.M. et al. Biotechnol Lett (2010) 32: 1117. doi:10.1007/s10529-010-0265-1

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.

Keywords

Phenylalanine production Proteolysis TyrA-tag Tyrosine by-production 

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Vera G. Doroshenko
    • 1
  • Rustem S. Shakulov
    • 2
  • Svetlana M. Kazakova
    • 1
  • Alexander D. Kivero
    • 1
  • Tatyana A. Yampolskaya
    • 1
  • Sergey V. Mashko
    • 1
  1. 1.Ajinomoto-Genetika Research InstituteMoscowRussian Federation
  2. 2.State Research Institute of Genetics and Selection of Industrial MicroorganismsMoscowRussian Federation

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