Comprehensive genome-scale and pathway engineering for production of the sustainable aviation fuel in Pseudomonas putida

Biological production of aviation fuels has a realistic impact on global warming. Important sustainable aviation fuel (SAF) targets are emerging and isoprenol is recently identified as a precursor for a promising SAF compound DMCO (1,4-dimethylcyclooctane). Isoprenol has been produced in several engineered microorganisms, and recently, Pseudomonas putida has gained interest as a promising host for isoprenol bioproduction as it can utilize carbon sources generated from inexpensive plant biomass. Here, we engineer metabolically versatile host P. putida for isoprenol production. We employ two computational modeling approaches (Bilevel optimization and Constrained Minimal Cut Sets) to predict gene knockout targets and optimize the “IPP-bypass” pathway in P. putida to maximize isoprenol production. Altogether, the highest isoprenol production titer from P. putida was achieved at 3.5 g/L under fed-batch conditions. This combination of computational and biological engineering on P. putida for an advanced biofuels production has vital significance in enabling a bioproduction process that can use renewable carbon streams.