Fig. 4

(a) HPLC chromatograms of cell extracts of SCMO1F, SCMO1F-3, and SCMO2-3 were analyzed. Peaks 1 and 2 are 3-dehydrorabelomycin and oviedomycin, respectively. (b) Results of LC/MS of 3-dehydrorabelomycin (m/z: 320, Up: 321.3 [M + H⁺]; bottom: 319.6 [M + H]). (c) Oviedomycin biosynthetic pathway. Refactoring of the target gene is indicated by the red arrow. Ovm BGC is shown with the corresponding pathway (yellow, oxygenase; gray, cyclase; green, polyketide synthase; orange, ketoreductase; blue, aromatase; and pink, 4’-phosphopantetheinyl transferase). (d) Analysis of oviedomycin production in SCMO2 with individual overexpression of the target genes, and also with overexpression of pairwise combinations of the target genes. The three target genes were predicted using the GEM. In the table, ‘O’ and ‘-’ denote the inclusion and absence of overexpressed gene(s), respectively. Data represent the average and standard deviation of three independent experiments