E. coli strains were cultivated in 2× yeast extract–tryptone medium supplemented with 100 µg/mL ampicillin, 100 µg/mL apramycin, or 50 µg/mL kanamycin, as required, and cultivated at 28 °C. MS medium agar and TSB (1.7 % casein peptone, 0.3 % soy peptone, 0.25 % glucose, 0.5 % sodium chloride, 0.25 % dipotassium hydrogen phosphate; [40]) were used with incubations at 28 °C for sporulation and for cultivation of S. rimosus strains in liquid medium, respectively. For OTC production, S. rimosus was cultivated in GOTC-V seed medium (5 % tryptone, 1 % glucose, 0.1 % calcium carbonate, 0.5 % yeast extract), incubated for 24 h at 28 °C with 220 rpm shaking. Then, 10 % (v/v) of the seed culture was used for inoculation of the GOTC-P production medium (0.7 % MOPS, 4.2 % soy flour, 0.6 % ammonium sulfate, 0.2 magnesium chloride, 0.15 % sodium chloride, 0.73 % calcium carbonate, 2.8 % corn starch, 10 mL/L 1 % zinc sulfate solution, 3.75 mL/L 1 % manganese sulfate solution, pH 6.25). The strains were cultivated in production medium for 5 days at 28 °C with 220 rpm shaking, and 60 % humidity. Intergeneric conjugation was performed according to a previously reported procedure [39]. S. rimosus exconjugants were selected on MS medium plates containing nalidixic acid (25 µg/mL) and thiostrepton (30 µg/mL), or erythromycin (30 µg/mL). After selection, the colonies were patched onto MS medium containing the appropriate antibiotic and incubated for 7 days at 28 °C.

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