Additional file 1 of Adaptive laboratory evolution accelerated glutarate production by Corynebacterium glutamicum

Additional file 1: Figure S1. (A) DNA region of the plasmid pEC-XT99A-gabTDP134L with the 21 bp sequence deleted in ALE strain GluA T7 (boxed in red). (B) Determination of the minimum inhibitory concentration for tetracycline for strain GluA T0 with the orginal plasmid pEC-XT99A-gabTDP134L and GluA RG1 carrying this plasmid with the 21 bp deletion. Cells were grown in Duetz cultivation system using BHIS supplemented with increasing concentrations of tetracycline (0–80 µg µL−1). The ΔOD600 values were determined after 48 hours and are shown as means with standard deviations (n = 3 cultivations). Figure S2. Dependence of the specific acitvity of GOGAT on the glutamine concentration determined for GRLys1ΔgltB overexpressing the native version of gltBD (A) or the mutated gltBE686QD (B). Crude extracts of GRLys1ΔgltB (pEKEx3-gltBD) and GRLys1ΔgltB (pEKEx3-gltBE686QD) were assayed for specific activity of the glutamate synthase (GOGAT) in the presence of increasing glutamine concentrations (0 mM, 0.1 mM, 0.25 mM, 0.5 mM, 0.75 mM, 1 mM, 2 mM, 5 mM, 10 mM). Values represent means and standard deviations of triplicate measurements. Figure S3. Glutarate production by GluA T7 in chloride-free medium. Cells were grown in Duetz cultivation system using either standard CGXII minimal medium or CGXII minimal medium without chloride supplemented with 40 g L−1 glucose, induced with 1 mM IPTG, and harvested after 48 h. Values and error bars represent means and standard deviations of glutarate (green) and 5AVA concentrations (red) in the culture supernatants (n = 3 cultivations).