D-Ribose plays fundamental roles in all living organisms and has been applied in food, cosmetics, health care, and pharmaceutical sectors. At present, D-ribose is predominantly produced by microbial fermentation based on the pentose phosphate pathway (PPP). However, this method suffers from a long synthetic pathway, severe growth defect of the host cell, and carbon catabolite repression (CCR). According to the Izumoring strategy, D-ribose can be produced from D-xylose through only three steps. Being not involved in the growth defect or CCR, this shortcut route is promising to produce D-ribose efficiently. However, this route has never been demonstrated in engineering practice, which hinders its application. In this study, we stepwise demonstrated this route and screened out higher active enzymes for each step. The first D-ribose production from D-xylose through the Izumoring route was achieved. By stepwise enzyme dosage tuning and process optimization, 6.87 g/L D-ribose was produced from 40 g/L D-xylose. Feeding D-xylose further improved the D-ribose titer to 9.55 g/L. Finally, we tested the coproduction of D-ribose and D-allose from corn stalk hydrolysate using the route engineered herein. In conclusion, this study demonstrated a pentose Izumoring route, complemented the engineering practices of the Izumoring strategy, paved the way to produce D-ribose from D-xylose, and provided an approach to comprehensively utilize the lignocellulosic sugars.

Producing D-Ribose from D-Xylose by Demonstrating a Pentose Izumoring Route