Fig. 6: Production of DLA from DLAM33B in 1 and 4 L scale fermentation.

The fermentation process was modeled after the induction protocol used in shake flask experiments, with additional galactose and 10X SC-URA media supplemented at defined feeding phases (I, II, III) in a fed-batch mode. 50 mM Ammonium-succinate was supplemented into the culture media from the onset to maintain a pH of 5.8. Both 1 and 4 L culture fermentations achieved a maximum wet cell mass of around 26 g L⁻¹ and a maximum DLA titer of 1.7 mg L⁻¹ and 1.4 mg L⁻¹. Feeding phase I: initial induction phase mimicking the addition of galactose for induction in shake flask experiments. 10X feed solutions were added to the vessel to a final concentration of 1X, at a rate of 3.5 ml min⁻¹. Feeding phase II: sustained feeding phase, a second round of 10X feed solutions were supplemented to the culture to a final concentration of 1X over a period of 52 h. Feeding phase III: starvation phase, no additional carbon or nitrogen source was supplemented. Data are presented as mean values + /− standard deviation. Error bars were calculated from three biological replicates. Source data are provided as a Source Data file.