Biosynthesis of suberic acid from glucose through the inverted fatty acid β-oxidation by recombinant Escherichia coli strains

Using directly engineered derivatives of previously constructed adipate-producingEscherichia colistrains MG1655lacIQ, ∆ackA-pta, ∆poxB, ∆ldhA,∆adhE, PL-SDj10-atoB, Ptrc-ideal-4-SDj10-fadB, ∆fadE, PL-SDj10-tesB, ∆yciA, Ptrc-ideal-4-SDj10-fabI, PL-SDj10-paaJ,∆aceBAK,∆glcBи MG1655lacIQ, ∆ackA-pta, ∆poxB, ∆ldhA,∆adhE, PL-SDj10-atoB, Ptrc-ideal-4-SDj10-fadB, PL-SDj10-tesB, ∆yciA, Ptrc-ideal-4-SDj10-fadE, PL-SDj10-paaJ,∆aceBAK,∆glcBthe feasibility of suberic acid biosynthesis from glucose by this bacterium resulting from the reversal of the native fatty acid β-oxidation pathway was demonstrated. The condensation of acetyl-CoA with succinyl-CoA and adipyl-CoA was ensured in recombinants by 3-oxoadipyl-CoA thiolase PaaJ, whereas the putative acetyl-CoA C-acetyltransferase YqeF was unable to catalyse the respective reactions.The biosynthesis of ~60 μM suberic acid was achieved upon significant enhancement in the strains of the expression of the bifunctional (S)-3-hydroxyacyl-CoA dehydrogenase/enoyl-CoA reductase gene,fadB. Subsequent inactivation of succinate dehydrogenase in the strains increased the intracellular availability of succinyl-CoA for the initiation of the first round of cycle reversal and favored an increase in the accumulation of the target compound by the recombinants to ~75 μM. The results provide a framework for the development of highly efficient producing strains for bio-based production of suberic acid from renewable raw materials.