Development of a Thermodynamically Favorable Multi‐enzyme Cascade Reaction for Efficient Sustainable Production of ω‐Amino Fatty Acids and α,ω‐Diamines

Aliphatic ω‐amino fatty acids (ω‐AFAs) and α,ω‐diamines (α,ω‐DMs) are essential monomers for the production of nylons. Development of a sustainable biosynthesis route for ω‐AFAs and α,ω‐DMs is crucial in addressing the challenges posed by climate change. Herein, we constructed an unprecedented thermodynamically favorable multi‐enzyme cascade (TherFavMEC) for the efficient sustainable biosynthesis of ω‐AFAs and α,ω‐DMs from cheap α,ω‐dicarboxylic acids (α,ω‐DAs). This TherFavMEC was developed by incorporating bioretrosynthesis analysis tools, reaction Gibbs free energy calculations, thermodynamic equilibrium shift strategies and cofactor (NADPH&ATP) regeneration systems. The molar yield of 6‐aminohexanoic acid (6‐ACA) from adipic acid (AA) was 92.3 %, while the molar yield from 6‐ACA to 1,6‐hexanediamine (1,6‐HMD) was 96.1 %, which were significantly higher than those of previously reported routes. Furthermore, the biosynthesis of ω‐AFAs and α,ω‐DMs from 20.0 mM α,ω‐DAs (C6‐C9) was also performed, giving 11.2 mM 1,6‐HMD (56.0 % yield), 14.8 mM 1,7‐heptanediamine (74.0 % yield), 17.4 mM 1,8‐octanediamine (87.0 % yield), and 19.7 mM 1,9‐nonanediamine (98.5 % yield), respectively. The titers of 1,9‐nonanediamine, 1,8‐octanediamine, 1,7‐heptanediamine and 1,6‐HMD were improved by 328‐fold, 1740‐fold, 87‐fold and 3.8‐fold compared to previous work. Therefore, this work holds great potential for the bioproduction of ω‐AFAs and α,ω‐DMs. Two thermodynamically favorable multi‐enzyme cascade (Module 1 and Module 2) were constructed for synthesis of important nylon monomers, ω‐AFAs and α,ω‐DMs from cheap and readily available bio‐based α,ω‐DAs, with the higest titers of 11.2–19.7 mM. The titers of 1,9‐nonanediamine,1,8‐octanediamine, 1,7‐heptanediamine and 1,6‐HMD were improved by 328‐fold, 1740‐fold, 87‐fold and 3.8‐fold compared to previous work.