Fig. 3

Semi-aerobic cultivation of T. aromatica producing S25DH₁ with nitrate as electron acceptor. (A), Representative growth curve of T. aromatica producing S25DH₁ with acetate + nitrate (1:1) under aerobic conditions without stirring/shaking at 30 °C (semi-aerobic, black line), and formation of 25OHVitD₃ from 500 µM VitD₃ (grey bars) by cells taken at different time points (0.5 mL scale, OD 50 corresponding to 8 mg cells [dry weight], 5% [w/v] HPCD, 1% [v/v] isopropanol, aerobic). (B), Representative growth curve under semi-aerobic conditions (black line) and oxygen consumption (blue squares). (C), Growth curve to maximal OD_{578 nm} and the effect of supplementation as indicated by the horizontal dashed lines. (D), Stability of T. aromatica cells producing S25DH₁. Cells were grown under aerobic conditions + nitrate at 30 °C. After consumption of acetate/nitrate, the resting cells were stored at 4 °C; OD_{578 nm} (black line); 25OHVitD₃ formation within 2 h from 500 µM VitD₃ (grey bars) by cells taken at different time points (0.5 mL scale, OD 100 corresponding to 16 mg cells [dry weight], 5% [w/v] HPCD, 1% [v/v] isopropanol, aerobic)