Fig. 3: Increasing CvFAP copy number debottlenecked the pathway for alka(e)ne production.

a Metabolic pathways for improving alka(e)ne production by increasing CvFAP copy number. Abbreviations: ACC, acetyl-CoA carboxylase; FAS1 and FAS2, fatty acid synthase 1 and 2; DGAs, diacylglycerol transferases; ALK1 and ALK2, cytochrome P450ALKs belonging to the CYP52 family with major functions in n-alkane assimilation;⁴⁶ FAA1 was found to be related to n-alkane assimilation¹⁹. b Overexpression of CvFAP in strains with double deletions of FAA1 and ALK2 and triple deletions of FAA1, ALK1, and ALK2 enhanced alka(e)ne synthesis more profoundly (n = 2 biologically independent samples). c Compared with the parental strain YLjbl-19, CvFAP, CvFAPD, and CvFAPT transformants showed 1.7, 2.4, and 2.4-fold higher alka(e)ne titers, respectively (n = 3 biologically independent samples). d Relative copy numbers of the CvFAP gene were quantified and showed a positive correlation with alka(e)ne titers (n = 3 biologically independent samples). e In addition, pentadecene (C15:1) derived from palmitoleoyl-CoA was detected in these strains but not the parental strains (observed from at least three biologically independent samples). f Alka(e)ne production of CvFAP, CvFAPD, and CvFAPT transformants from YLjbl-20 was improved as well, but not as much as the transformants from YLjbl-19 (n = 3 biologically independent samples). Fermentations in (b) were performed in 50 mL glass conical shake flasks with a working volume of 13 mL and an initial OD₆₀₀ of 0.1. Samples were collected after 3 days of cultivation in the dark followed by 3 days in blue light generated by light source 1 (Supplementary Fig. 16). Other fermentations were performed in 12-well microplates with a working volume of 2 mL and an initial OD₆₀₀ of 0.5. Samples were collected after 2 days of cultivation in the dark at 30 °C followed by 1 day of cultivation in blue light generated by light source 2 at 25 °C (Supplementary Fig. 16). Fermentation medium was composed of 20 g/L glucose, 6.9 g/L yeast nitrogen base (without amino acids), and 1 g/L yeast extract. Data represent mean value ± SD. Source data underlying Fig. 3b–d, f are provided as a Source Data file.