Efficient Enzymatic Cascade Synthesis of α-Olefins via Colocalization onto Graphene Oxides and Liquid–Liquid Extraction

α-Olefins are important precursors for many chemical products such as lubricants and detergents. The biocatalytic reaction has become a new approach to synthesizing α-olefins from biobased feedstocks such as fatty acids. Two enzymes, pyranose-2-oxidase (P2O) and cytochrome P450-OleTJE (OleTJE), are coupled to form a cascade reaction in which the first P2O-catalyzed reaction generates H2O2, which is a cosubstrate for the second OleTJE-catalyzed reaction to convert fatty acids into α-olefins. In this work, we developed multienzyme immobilization protocols that colocalize the two enzymes onto graphene oxide (GO) in close proximity, enabling efficient diffusion of H2O2 between the reactions. The coimmobilized enzymes on GO exhibited enhanced productivity with a relative activity of 143% as compared to that of the free enzymes. Also, liquid–liquid extraction (LLE) was used to recover α-olefins and facilitate enzyme reuse. When employing LLE with dodecane as a solvent, the overall production increased by 2.47-fold compared to the conventional condition of using free enzymes with ethyl acetate as a solvent.