To quantify the OTC titers from the S. rimosus strains, 1 mL of fermentation broth was adjusted to pH 1.5-2.0 with 37 % HCl, and centrifuged at 13,000 rpm for 10 min. The supernatants were then subjected to HPLC analysis (UltiMate 3000 HPLC system) with a C-18 column (150 × 4.6 mm; 5 µm; 40 °C; Macherey-Nagel), with UV absorption detection at 270 nm, and a flow rate of 0.3 mL/min. The elution solvents were water with 0.1 % formic acid (solvent A) and acetonitrile (solvent B). The 30-min gradient elution was as follows: 0→20 min, 5 %→25 % B; 20→21 min, 25 %→90 % B; 21→25 min, 90 % B; 25→26 min, 90 %→5 % B; 26→30 min, 5 % B. The OTC titers were calculated as the mean titers from three parallel samples of each transformant and parental strain.

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