For flavonoid production from L-tyrosine, strains were first cultured in 25 mL of MOPS medium at 37°C with 220 rpm orbital shaking. After an OD600 of 1.65 had been reached, an additional 25 mL of fresh MOPS medium, a final aliquot of isopropyl-β-D-thiogalactopyranoside (IPTG) (taking the concentration to 1 mM), and a final aliquot of L-tyrosine (taking the concentration to 3 mM) were added. Cultures were subsequently conducted at 30°C for (2S)-naringenin production. (2S)-Naringenin concentrations were measured after a total fermentation time of 48 h. For malonyl-CoA availability experiments, 1 g/L of sodium malonate dibasic (Sigma) was added twice, resulting in a total concentration of 2 g/L.
For flavonoid production from D-glucose, strains were first cultured in 25 mL of MOPS medium until an OD600 of 1.65 was reached, after which an additional 25 mL of fresh MOPS medium and a final aliquot of IPTG (taking the concentration to 1 mM) were added. Cultures were subsequently conducted at 30°C for (2S)-naringenin production. For malonyl-CoA availability experiments, sodium malonate dibasic (Sigma) was added at a concentrations of 2 g/L (1 g/L added twice).
For flavonoid production from D-glucose, strains were first cultured in 25 mL of MOPS medium until an OD600 of 1.65 was reached, after which an additional 25 mL of fresh MOPS medium and a final aliquot of IPTG (taking the concentration to 1 mM) were added. Cultures were subsequently conducted at 30°C for (2S)-naringenin production. For malonyl-CoA availability experiments, sodium malonate dibasic (Sigma) was added at a concentrations of 2 g/L (1 g/L added twice).