Figure 4

dCyd production by all engineered BL21 derivative strains. Seed cultures were grown in 5 mL of Luria–Bertani media overnight, and then transferred to 500-mL baffled flasks containing 50 mL of production media (see “Methods”). The strains carrying expression plasmids were induced for protein expression with isopropyl β-d-1-thiogalactopyranoside (IPTG; 0.5 mM) when cells reached OD₆₀₀ ≈ 0.6. All flask cultures were incubated at 37°C and 250 rpm for 36 h. The dCyd concentration in the media was determined from the supernatants of cultivation samples. Values are mean ± SD of triplicate flasks experiments performed at the same time.