Zha, Jian Zang, Ying Mattozzi, Matthew Plassmeier, Jens Gupta, Mamta Wu, Xia Clarkson, Sonya Koffas, Mattheos MOESM1 of Metabolic engineering of Corynebacterium glutamicum for anthocyanin production Additional file 1: Figure S1. SDS-PAGE (10% gel) analysis of proteins ANS and 3GT by recombinant C. glutamicum expressing ANS and 3GT in various media. The E. coli strain expressing ANS and 3GT was used as a positive control. E. coli was cultivated in AMM with 2% glucose and induced by 1 mM IPTG for 4 h before harvested for protein extraction. The C. glutamicum strain was grown in BHIS, AMM or CGXII medium and was induced by 1 mM IPTG at mid-exponential phase for 12 h. Red arrows indicate bands for ANS (48.5 kD) and 3GT (50.5 kD) (their molecular weight is too close, and only one merged band could be seen in the gel). Figure S2. Mass Spectrum identification of C3G in the fermentation products of recombinant C. glutamicum strains. TIC (A) and EIC (B) for mass range of C3G of the standard C3G; TIC (C) and EIC (D)for mass range of C3G for the fermentation products; Mass spectrum for C3G peak in the standard (E) and the fermentation products (F). Figure S3. Time course study of mCherry expression in C. glutamicum. The mCherry gene was cloned into the plasmid pZM1, and the expression of mCherry was indicated by the fluorescence intensity at an excitation wavelength of 588 nm and an emission wavelength of 618 nm. Table S1. Plasmids and strains used in the present study. Table S2. Primers used in this study. Table S3. C3G production using concentrated cells in different conditions. Cells grown in AMM (pH7.0) were induced with IPTG (0.5 mM) for 6 h and harvested. Then 5 ml of cells were resuspended in 1 ml of different buffers with 500 mg/L catechin and necessary supplements, including citrate buffer, potassium phosphate buffer, AMM (pH 5.0) and AMM (pH 7.0), respectively. The conversion process was conducted at 30 °C and 220 rpm for 48 h. Anthocyanin;Corynebacterium glutamicum;Catechin;Flavonoid;UDP-glucose 2018-09-14
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