title: Metabolic Engineering of Pseudomonas putida KT2440 to Produce Anthranilate from Glucose.
abstract: The Pseudomonas putida KT2440 strain was engineered in order to produce anthranilate (oAB, ortho-aminobenzoate), a precursor of the aromatic amino acid tryptophan, from glucose as sole carbon source. To enable the production of the metabolic intermediate oAB, the trpDC operon encoding an anthranilate phosphoribosyltransferase (TrpD) and an indole-3-glycerol phosphate synthase (TrpC), were deleted. In addition, the chorismate mutase (pheA) responsible for the conversion of chorismate over prephenate to phenylpyruvate was deleted in the background of the deletion of trpDC to circumvent a potential drain of precursor. To further increase the oAB production, a feedback insensitive version of 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase encoded by the aroG (D146N) gene and an anthranilate synthase (trpE (S40F) G) were overexpressed separately and simultaneously in the deletion mutants. With optimized production conditions in a tryptophan-limited fed-batch process a maximum of 1.54 ± 0.3 g L(-1) (11.23 mM) oAB was obtained with the best performing engineered P. putida KT2440 strain (P. putida ΔtrpDC pSEVA234_aroG (D146N) _trpE (S40F) G).
authors.affiliation: Institute of Applied Microbiology (iAMB), Aachen Biology and Biotechnology (ABBt), Rheinisch-Westfälische Technische Hochschule Aachen University Aachen, Germany.
authors.forename: Jannis
authors.initials: J
authors.lastname: Kuepper
authors.affiliation: Institute of Applied Microbiology (iAMB), Aachen Biology and Biotechnology (ABBt), Rheinisch-Westfälische Technische Hochschule Aachen University Aachen, Germany.
authors.forename: Jasmin
authors.initials: J
authors.lastname: Dickler
authors.affiliation: Institute of Applied Microbiology (iAMB), Aachen Biology and Biotechnology (ABBt), Rheinisch-Westfälische Technische Hochschule Aachen University Aachen, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Biggel
authors.affiliation: Bayer Technology Services GmbH Leverkusen, Germany.
authors.forename: Swantje
authors.initials: S
authors.lastname: Behnken
authors.affiliation: Bayer MaterialScience AG Leverkusen, Germany.
authors.forename: Gernot
authors.initials: G
authors.lastname: Jäger
authors.affiliation: Institute of Applied Microbiology (iAMB), Aachen Biology and Biotechnology (ABBt), Rheinisch-Westfälische Technische Hochschule Aachen University Aachen, Germany.
authors.forename: Nick
authors.initials: N
authors.lastname: Wierckx
authors.affiliation: Institute of Applied Microbiology (iAMB), Aachen Biology and Biotechnology (ABBt), Rheinisch-Westfälische Technische Hochschule Aachen University Aachen, Germany.
authors.forename: Lars M
authors.initials: LM
authors.lastname: Blank
keywords: Pseudomonas putida KT2440; anthranilic acid; aromatic amino acid pathway; industrial biotechnology; metabolic engineering