Metabolic engineering of the E. coli L-phenylalanine pathway for the production of D-phenylglycine (D-Phg).
D-phenylglycine (D-Phg) is an important side chain building block for semi-synthetic penicillins and cephalosporins such as ampicillin and cephalexin. To produce d-Phg ultimately from glucose, metabolic engineering was applied. Starting from phenylpyruvate, which is the direct precursor of L-phenylalanine, an artificial D-Phg biosynthesis pathway was created. This three-step route is composed of the enzymes hydroxymandelate synthase (HmaS), hydroxymandelate oxidase (Hmo), and the stereoinverting hydroxyphenylglycine aminotransferase (HpgAT). Together they catalyse the conversion of phenylpyruvate via mandelate and phenylglyoxylate to D-Phg. The corresponding genes were obtained from Amycolatopsis orientalis, Streptomyces coelicolor, and Pseudomonas putida. Combined expression of these activities in E. coli strains optimized for the production of L-phenylalanine resulted in the first completely fermentative production of D-Phg.
Müller
Ulrike
U
DSM Biotech GmbH, Karl-Heinz-Beckurts-Str. 13, 52428 Jülich, Germany. ulrike.mueller@dsm.com
van Assema
Friso
F
Gunsior
Michele
M
Orf
Sonja
S
Kremer
Susanne
S
Schipper
Dick
D
Wagemans
Anja
A
Townsend
Craig A
CA
Sonke
Theo
T
Bovenberg
Roel
R
Wubbolts
Marcel
M
D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D024861:Genetic Enhancement / Q000379:methods; D005947:Glucose / Q000378:metabolism*; D005998:Glycine / Q000031:analogs & derivatives* / Q000235:genetics / Q000378:metabolism; D009097:Multienzyme Complexes / Q000235:genetics / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism
D009097:Multienzyme Complexes; D011994:Recombinant Proteins; C008852:2-phenylglycine; D005947:Glucose; D005998:Glycine