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Xylose reductase (XR), an industrially important enzyme, catalyzes the hydrogenation of xylose into xylitol. Xylitol, a polyol sugar, has tremendous applications in different industries due to its significant properties. XR is mainly produced from yeasts and molds, and limited studies have been reported on bacteria. The present study explores the potential of newly isolated bacteria, i.e., Pseudomonas putida BSX-46 for XR synthesis through process scale-up by tailoring its nutritional and cultural requirements. A simple media containing only four ingredients was designed for the production of XR in a short incubation time of 24 h. A process for pretreatment of rice straw was developed to achieve hydrolysate with a good amount of xylose (140 g/kg of rice straw). The enhanced XR production of 213.14±0.47 IU/mg of cells was achieved at bioreactor level using waste rice straw hydrolysate as compared to 94.26±0 …