What to sacrifice? Fusions of cofactor regenerating enzymes with Baeyer-Villiger monooxygenases and alcohol dehydrogenases for self-sufficient redox biocatalysis

Abstract A collection of fusion biocatalysts has been generated that can be used for self-sufficient oxygenations or ketone reductions. These biocatalysts were created by fusing a Baeyer-Villiger monooxygenase (cyclohexanone monooxygenase from Thermocrispum municipale : Tm CHMO) or an alcohol dehydrogenase (alcohol dehydrogenase from Lactobacillus brevis : Lb ADH) with three different cofactor regeneration enzymes (formate dehydrogenase from Burkholderia stabilis : Bs FDH; glucose dehydrogenase from Sulfolobus tokodaii : St GDH, and phosphite dehydrogenase from Pseudomonas stutzeri : Ps PTDH). Their tolerance against various organic solvents, including a deep eutectic solvent, and their activity and selectivity with a variety of substrates have been studied. Excellent conversions and enantioselectivities were obtained, demonstrating that these engineered fusion enzymes can be used as biocatalysts for the synthesis of (chiral) valuable compounds.