MOESM1 of The two-component system CepRS regulates the cephamycin C biosynthesis in Streptomyces clavuligerus F613-1

Additional file 1: Table S1. Plasmids and strains used in this study. Table S2. Primers used in this study. Figure S1. (A) Polymerase chain reaction-based verification of the ΔcepS and ΔcepR strains. M: DNA marker. (B) Phenotype of F613-1, △cepRS, △cepR, △cepS and △cepRS on BSCA solid media. (C) Ceph C (cephamycin C) obtained in F613-1, △cepRS, △cepR and △cepS strains on TSA solid medium. *, statistically significant difference (P < 0.05) between △cepRS, △cepR, △cepS and F613-1 at the same time point. Figure S2. Biomass and clavulanic acid (CA) concentration of F613-1, △cepRS and the complemented strain. A: Growth curves of F613-1, △cepRS and △cepRScom in SCF fermentation medium. Samples for growth curve analysis were harvested at five time points (1, 3, 5, 7 and 9 d). Data are the mean ± SD of three independent experiments. B: Analysis of the change of CA concentration during fermentation. Data are the mean ± SD of three independent biological experiments. Figure S3. Expression of genes in the CA biosynthetic gene cluster were examined by RT-qPCR between F613-1 (black bars), △cepRS (grey bars) and △cepRScom (dark grey bars). Results were normalized for 16S rRNA gene content and are shown as fold change over the F613-1 control. Data are the mean ± SD of three independent biological experiments. Figure S4. A: Phenotype of F613-1, △cepRS, △cepRScom and △cepRS-cepRS (over-expression strain) on BSCA solid media. B: Cephamycin C obtained in F613-1, △cepRS and △cepRS-cepRS (over-expression strain) strains on TSA solid medium. *, compared with F 613-1, P < 0.05. Data are the mean ± SD of three independent biological experiments. C: Expression of genes in the cephamycin C biosynthetic gene cluster were examined by RT-qPCR between F613-1 (black bars), △cepRS (grey bars) and △cepRS-cepRS (over-expression strain) (dark grey bars). Results were normalized for 16S rRNA gene content and are shown as fold change over the F613-1 control. *, compared with F613-1, P < 0.05. Data are the mean ± SD of three independent biological experiments.