Efficient production of inositol from glucose via a tri-enzymatic cascade pathway

Inositol is an essential intermediate in cosmetics, food, medicine and other industries. However, developing an efficient biotransformation system for large-scale production of inositol remains challenging. Herein, a tri-enzymatic cascade route with three novel enzymes including polyphosphate glucokinase (PPGK) from Thermobifida fusca, inositol 3-phosphate synthase (IPS) from Archaeoglobus profundus DSM 5631 and inositol monophosphatase (IMP) from Thermotoga petrophila RKU-1 was designed and reconstructed for the production of inositol from glucose. The problem of poor cooperativity of the cascade reactions was addressed by ribosome binding site (RBS) optimization of PPGK and replication of IPS. Under the optimum biotransformation conditions, the engineered whole-cell immobilized with colloidal chitin transformed 120 g/L glucose to 110.8 g/L inositol with 92.3% conversion in four cycles of reuse, representing the highest titer of inositol to date. Furthermore, this is the first study for inositol production using a three-enzyme coordinated immobilized single-cell.