Fig. 4: Redirection of acyl-CoA to form alka(e)nes during fed-batch fermentation process.

a Production of alka(e)nes by YLjbl-26 in fed-batch fermentations. Gray-colored regions indicate cultivation in the dark while blue-colored regions indicate cultivation in blue light (n = 3 or 4 biologically independent samples). b Alka(e)ne titers and productivities from different glucose loadings with the same feeding strategy as (a) (n = 2 biologically independent samples). c The amount of lipids in cells cultured in dark conditions matched the sum of lipids and alka(e)nes in cells cultured in light conditions. Both fermentations were carried out following the same feeding strategy as shown in (a). d We further developed a method (Supplementary Note 2) to determine the fraction of the redirected flux. This fraction increased from 31% to 85% as fermentation progressed from day 3 to 11, asymptotically reaching a maximum of 89%. e Brightfield microscopic images of cells with alka(e)ne titers of 1.03 g/L, equivalent to 89% of neutral lipids. f Fluorescence images of cells (yellow), merged with the brightfield images. Staining our alka(e)ne-producing strain reveals that most cells were filled with intracellular lipids and alka(e)nes with no evidence of extracellular fluorescence. Images were taken by a DeltaVision2-TIRF Microscope. Cells were stained by Nile red. Fermentations were performed in 50 mL glass conical shake flasks with a working volume of 13 mL and an initial OD₆₀₀ of 0.1. Blue light was generated by light source 2 (Supplementary Fig. 16). Data represent mean value ± SD, n = 3 or n = 4 biologically independent samples. Source data underlying Fig. 4a–d are provided as a Source Data file.