Rational design of 3-D electroenzymatic electrodes comprising novel enzymatic cascade and NADP(H) co-factor regeneration

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Electroenzymatic processes have enormous potential for the development of new biotechnological processes for fine and commodity chemicals, but there are some major bottlenecks, like product scope, co-factor regeneration and productivity which impede their technical applications. In this project these issues are addressed by joint efforts between the biocatalysis (Bornscheuer) and electrochemical engineering (Vidakovic-Koch) groups. A model enzymatic cascade which exemplifies some typical problems to be faced by the integration of enzymatic catalysts and electrochemical regeneration will be studied. It comprises the combination of enoate reductases (ERED) and Baeyer-Villiger monooxygenases (BVMO). Both enzyme types require the soluble NAD(P)H co-factor which is a versatile co-factor required for a broad range of enzymatic reduction reactions of significant importance for global pharmaceutical market. We aim for a direct NAD(P)H regeneration at the electrode surface, without any further components, except electrons, involved. To combine the co-factor regeneration with a monooxygenase / enoate reductase enzymatic cascade, novel 3-D porous electrodes will be developed. The macro kinetics of the enzymatic cascade reaction with co-factor regeneration will be assessed in model-based analysis. Rational protein design and directed evolution will be used to adapt the enzymes to conditions required for efficient electroenzymatic reactor.