Construction of an l-phenylalanine-producing tyrosine-prototrophic Escherichia coli strain using tyrA ssrA-like tagged alleles |
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Authors: | Vera G. Doroshenko Rustem S. Shakulov Svetlana M. Kazakova Alexander D. Kivero Tatyana A. Yampolskaya Sergey V. Mashko |
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Affiliation: | 1. Ajinomoto-Genetika Research Institute, Moscow, 117545, Russian Federation 2. State Research Institute of Genetics and Selection of Industrial Microorganisms, Moscow, 117545, Russian Federation
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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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