Identification, Characterization, and Application of Three Enoate Reductases from Pseudomonas putida in In Vitro Enzyme Cascade Reactions

Enoate reductases are versatile enzymes for the enantio‐ and regioselective addition of hydrogen to double bonds. We identified three EREDs (XenA, XenB, NemA) from Pseudomonas putida ATCC 17453 through a sequence motif search. In addition to cloning, functional expression, and biochemical characterization of these enzymes, the enoate reductases were also applied in enzyme cascade reactions in combination with a Baeyer–Villiger monooxygenase and an alcohol dehydrogenase to produce lactones. Good things come in threes: The identification, cloning, expression, and characterization of three enoate reductases from Pseudomonas putida reveal broad substrate scope and high stereoselectivities. Furthermore, the enoate reductases could be integrated into cascade reactions together with an alcohol dehydrogenase and a Baeyer–Villiger monooxygenase.