Chitin, a widely available biomass, was efficiently converted to glucose using a modular multienzyme cascade without requiring ATP. By integrating known metabolic pathways, we designed and assembled a self-sustaining catalytic network comprising seven key enzymes. A key advance in this chitin-to-glucose transformation was the use of chitobiose phosphorylase, which utilizes inorganic phosphate as the phosphoryl donor, enabling the formation of phosphorylated sugar intermediates without requiring ATP. These findings provide new possibilities for the development of next-generation biorefineries utilising production of glucose from marine and agricultural chitinous residues.
