title: Metabolic engineering of the E. coli L-phenylalanine pathway for the production of D-phenylglycine (D-Phg).
abstract: 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.
authors.affiliation: DSM Biotech GmbH, Karl-Heinz-Beckurts-Str. 13, 52428 Jülich, Germany. ulrike.mueller@dsm.com
authors.forename: Ulrike
authors.initials: U
authors.lastname: Müller
authors.forename: Friso
authors.initials: F
authors.lastname: van Assema
authors.forename: Michele
authors.initials: M
authors.lastname: Gunsior
authors.forename: Sonja
authors.initials: S
authors.lastname: Orf
authors.forename: Susanne
authors.initials: S
authors.lastname: Kremer
authors.forename: Dick
authors.initials: D
authors.lastname: Schipper
authors.forename: Anja
authors.initials: A
authors.lastname: Wagemans
authors.forename: Craig A
authors.initials: CA
authors.lastname: Townsend
authors.forename: Theo
authors.initials: T
authors.lastname: Sonke
authors.forename: Roel
authors.initials: R
authors.lastname: Bovenberg
authors.forename: Marcel
authors.initials: M
authors.lastname: Wubbolts
mesh_terms: 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
chemical_list: D009097:Multienzyme Complexes; D011994:Recombinant Proteins; C008852:2-phenylglycine; D005947:Glucose; D005998:Glycine