Fig. 1

Engineering S. cerevisiae for enhanced fatty alcohol production via “spatial–temporal” regulation. In the early stage of fermentation (“the sun”), fatty acid biosynthesis was active for cell growth and a cytoplasmic fatty acyl-CoA reductase from Marinobacter sp. ES-1 (MaFAR1) was expressed under the control of strong constitutive promoters (orange arrow), coupled with a disrupted hexadecenal dehydrogenase (hfd1Δ). With the consumption of glucose (“the moon”), fatty acid/acyl-CoA was accumulated, which activated peroxisomal MaFAR1 by fatty acid/acyl-CoA responsive promoters (yellow arrow). The acyl-CoA transporter PXA1/PXA2 and a peroxisomal copy of acyl-CoA synthase FAA2 were overexpressed to increase peroxisomal acyl-CoA supply. The peroxisomal malate shuttling pathway (malic enzyme RtME, pyruvate carboxylase PYC1 and malate dehydrogenase MDH3), and isocitrate dehydrogenase (IDP2 and IDP3) were also targeted to peroxisomes to enhance the supply of peroxisomal NADPH. Peroxins (PEX7 and PEX28) were overexpressed and H₂O₂ was added in culture to promote peroxisome biogenesis. Overall, the coordination of fatty alcohol biosynthesis, precursor and cofactor supply, and peroxisome biogenesis was implemented for spatial–temporal regulation of fatty alcohol biosynthesis