Enzyme cascades for high-yield conversion of d-xylose into d-ribose by overcoming equilibrium constraints and...

d-Ribose is essential for critical cellular functions and the synthesis of antiviral nucleosides. However, traditional chemical synthesis and fermentation methods of d-ribose production suffer from low yields and inefficient resource utilization. Here, we present a highly efficient enzymatic cascade strategy that utilizes selective phosphorylation and dephosphorylation processes, coupled with ATP regeneration to convert d-xylose into d-ribose with high yield. By optimizing this enzyme cascade, we achieved a substantial increase in d-ribose yield from 23.4 % to 93.5 % mol/mol, effectively overcoming the equilibrium limitations of sugar conversion processes. Notably, our approach allows for the selective conversion of d-xylose to d-ribose in lignocellulosic hydrolysates, even in the presence of d-glucose. This work demonstrates the highly efficient enzymatic conversion of d-xylose into d-ribose offering a competitive alternative to existing chemical synthesis methods. Our findings provide a novel approach to cellulosic biomass valorization and represent a significant contribution to the field of biorefinery.