Figure 1: Engineering E. coli fatty acid metabolic pathway.

(a) E. coli fatty acid biosynthetic pathway. Overexpression targets are shown in green and knockout targets in red. Dash arrows represent multiple reaction steps. accABCD, acetyl-CoA carboxylase; fabD, malonyl-CoA-ACP transacylase; fabH, β-ketoacyl-ACP synthase III; fabB, β-ketoacyl-ACP synthase I; fabF, β-ketoacyl-ACP synthase II; fabG, β-ketoacyl-ACP reductase; fabA, β-hydroxy acyl-ACP dehydrase;fabI, enol acyl-ACP reductase; tesA′, truncated multifunctional fatty acyl-CoA thioesterase; and fadD, fatty acyl-CoA synthetase. (b) Fatty acid production in E. coli BL21ΔfadD expressing four subunits of acetyl-CoA carboxylase and plant fatty acyl-ACP thioesterase quantified by GC–MS and Nile red assay. Bars show fatty acid production analysed by GC–MS; red star shows fatty acid production analysed by Nile red assay. BnFatA, B. napus fatty acyl-ACP thioesterase (Gene ID: X73850.1); CnFatB2, C. nucifera fatty acyl-ACP thioesterase (Gene ID: JF338904.1); EGTE, E. guineensis fatty acyl-ACP thioesterase (Gene ID: AF110462.1). All experiments were performed in triplicate and error bars show s.d. with 95% CI. (c) High-performance liquid chromatography (HPLC) profile of fatty acids phenacyl esters from engineered strains expressing different thioesterases.