Supplemental methods (calcimycin production time course experiments, quantification of calG mRNA present in S. chartreusis NRRL 3882 WT cultures by qPCR); physicochemical data relating to compound 3, cezomycin, N-demethyl-calcimycin, calcimycin, and related SNAC and CoA variants (Table S1); Q-TOF MS characterization of compound 3, cezomycin, N-demethyl-calcimycin and calcimycin-related SNAC (Fig. S1) and CoA (Fig. S2) derivatives; protein sequence alignments of CalG with other typical TEIIs (Fig. S3); construction of the ΔcalG strain by PCR targeting (Fig. S4); HPLC assay of calcimycin and cezomycin production by S. chartreusis NRRL 3882, GLX 16, and GLX 17 strains grown in SFM liquid medium (Table S2); HPLC analysis of CalG SNAC derivative (Fig. S5) and CalG CoA derivative (Fig. S6) substrate usage; saturation curves for CalG with cezymycin-CoA and other CoA derivatives (Fig. S7); CalG substrate usage kinetics (Table S3); effect of divalent cations on production of calcimycin and cezomycin by S. chartreusis NRRL 3882 WT in SFM liquid medium (Table S4); calcimycin production by S. chartreusis NRRL 3882 WT cultured in SFM liquid medium over 9 days (Fig. S8); calG transcript profiling (Fig. S9); initial velocities of CalG hydrolysis of acetyl-CoA in the presence of divalent cations (Fig. S10).
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