FIG 1.

A novel synthetic pathway for glutarate production in E. coli. The reverse adipate degradation pathway (RADP) includes five steps in Thermobifida fusca: β-ketothiolase (Tfu_0875), 3-hydroxyacyl-CoA dehydrogenase (Tfu_2399), 3-hydroxyadipyl-CoA dehydrogenase (Tfu_0067), 5-carboxy-2-pentenoyl-CoA reductase (Tfu_1647), and adipyl-CoA synthetase (Tfu_2576-7). The arcA, ldhA, atoB, sucD, and pflB genes were deleted using the CRISPR/Cas9 system. A rough arrow represents genes or enzymes subject to overexpression, “X” represents those deleted, and the “⊥” symbol represents suppression. Solid arrows indicate single steps, three arrows indicate multiple steps, and dashed arrows indicate that a step does not exist in E. coli. ldhA, l-lactate dehydrogenase; atoB, acetyl-CoA C-acetyltransferase; arcA, aerobic respiration control protein; pflB, formate C-acetyltransferase 1; TCA, tricarboxylic acid cycle; EMP, the glycolysis pathway; OAA, oxaloacetate; PEP, phosphoenolpyruvate; PYR, pyruvate; SuccCoA, succinyl-CoA; ACP, acyl carrier protein.