AUTHOR=Kuepper Jannis , Dickler Jasmin , Biggel Michael , Behnken Swantje , Jäger Gernot , Wierckx Nick , Blank Lars M. 

TITLE=Metabolic Engineering of Pseudomonas putida KT2440 to Produce Anthranilate from Glucose

JOURNAL=Frontiers in Microbiology

VOLUME=6

YEAR=2015

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2015.01310

DOI=10.3389/fmicb.2015.01310

ISSN=1664-302X

ABSTRACT=<p>The <italic>Pseudomonas putida</italic> KT2440 strain was engineered in order to produce anthranilate (oAB, <italic>ortho</italic>-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 <italic>trpDC</italic> operon encoding an anthranilate phosphoribosyltransferase (TrpD) and an indole-3-glycerol phosphate synthase (TrpC), were deleted. In addition, the chorismate mutase (<italic>pheA</italic>) responsible for the conversion of chorismate over prephenate to phenylpyruvate was deleted in the background of the deletion of <italic>trpDC</italic> 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 <italic>aro</italic>G<sup><italic>D146N</italic></sup> gene and an anthranilate synthase (<italic>trpE<sup><italic>S40F</italic></sup>G</italic>) 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<sup>-1</sup> (11.23 mM) oAB was obtained with the best performing engineered <italic>P. putida</italic> KT2440 strain (<italic>P. putida</italic> Δ<italic>trpDC</italic> pSEVA234_<italic>aro</italic>G<sup><italic>D146N</italic></sup>_<italic>trpE<sup><italic>S40F</italic></sup>G</italic>).</p>