Division of Biofunctional Chemistry, The Chemical Society of Japan, Biotechnology Subcommittee

A dual-enzyme cascade, xylitol dehydrogenase and xylulose kinase derived from the xylose metabolic pathway, was constructed on a three-dimensional DNA scaffold with the dynamic shape transition from its open state to a closed state of a hexagonal prism. Evaluation of the cascade reaction efficiencies in the open and the closed states revealed little or no inter-enzyme distance dependence presumably due to the far larger catalytic constant of the downstream enzyme. The inter-enzyme distance is not the dominant factor for the cascade efficiency when kinetic parameters of the cascade enzymes are imbalanced with the highly efficient downstream enzyme.