Construction of an L-phenylalanine-producing tyrosine-prototrophic Escherichia coli strain using tyrA ssrA-like tagged alleles.
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.
Doroshenko
Vera G
VG
Ajinomoto-Genetika Research Institute, Moscow 117545, Russian Federation. vera_doroshenko@agri.ru
Shakulov
Rustem S
RS
Kazakova
Svetlana M
SM
Kivero
Alexander D
AD
Yampolskaya
Tatyana A
TA
Mashko
Sergey V
SV
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D008969:Molecular Sequence Data; D010649:Phenylalanine / Q000032:analysis / Q000096:biosynthesis*; D011303:Prephenate Dehydrogenase / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014443:Tyrosine / Q000032:analysis / Q000235:genetics*
D014443:Tyrosine; D010649:Phenylalanine; D011303:Prephenate Dehydrogenase