Production of tyrosine from sucrose or glucose achieved by rapid genetic changes to phenylalanine-producing Escherichia coli strains.
Escherichia coli K12 strains producing L-phenylalanine were converted to L-tyrosine-producing strains using a novel genetic method for gene replacement. We deleted a region of the E. coli K12 chromosome including the pheA gene encoding chorismate mutase/prephenate dehydratase, its leader peptide (pheL), and its promoter using a new polymerase chain reaction-based method that does not leave a chromosomal scar. For high level expression of tyrA, encoding chorismate mutase/prephenate dehydrogenase, its native promoter was replaced with the strong trc promoter. The linked DeltapheLA and Ptrc-tyrA::Kan(R) genetic modifications were moved into L-phenylalanine producing strains by generalized transduction to convert L-phenylalanine-producing strains to L-tyrosine-producing strains. Moreover, introduction of a plasmid carrying genes responsible for sucrose degradation into these strains enabled L-tyrosine-production from sucrose.
Olson
Monica M
MM
Biochemical Sciences and Engineering, DuPont Central Research and Development, Experimental Station, Wilmington, DE 19880, USA.
Templeton
Lori J
LJ
Suh
Wonchul
W
Youderian
Philip
P
Sariaslani
F Sima
FS
Gatenby
Anthony A
AA
Van Dyk
Tina K
TK
D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005947:Glucose / Q000378:metabolism*; D010649:Phenylalanine / Q000096:biosynthesis*; D013395:Sucrose / Q000378:metabolism*; D014443:Tyrosine / Q000096:biosynthesis*
D014443:Tyrosine; D010649:Phenylalanine; D013395:Sucrose; D005947:Glucose