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Biotechnol Lett ; 32(8): 1117-21, 2010 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-20364292

RESUMO

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 DeltatyrA. 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 TyrA(wt) strain.


Assuntos
Escherichia coli/genética , Escherichia coli/metabolismo , Fenilalanina/biossíntese , Tirosina/genética , Sequência de Aminoácidos , Cromatografia Líquida de Alta Pressão , Clonagem Molecular , Escherichia coli/crescimento & desenvolvimento , Dados de Sequência Molecular , Fenilalanina/análise , Prefenato Desidrogenase/genética , Prefenato Desidrogenase/metabolismo , Fatores de Tempo , Tirosina/análise
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