A hybrid system integrating xylose dehydrogenase and NAD

The wide application of xylose in the food, beverage, and pharmaceutical industries, as well as in the booming field of biorefinery, raises the demand for a rapid, accurate, and real-time xylose-sensing technique to rival the conventional methods based on chromatography, spectroscopy, and electrochemical analysis using non-specific enzymes or abiotic catalysts. Herein, a hybrid system comprising polyethylene glycerol swing-arm-tethered NAD+ and xylose dehydrogenase (XDH), coupled with platinum nanoparticles deposited on carbon nanotubes (PtNPs@MWCNTs), was constructed for the real-time sensing of xylose. The use of the PtNPs@MWCNTs composite enhanced the sensitivity of the electric response and reduced the oxidation potential of NADH significantly. Further, the NAD+ immobilization allowed an increase in its microenvironment concentration and facilitated cofactor regeneration. The screen-printed electrode cast with the hybrid system showed a wide xylose detection range of 0.5 to 10 mM or 3.33 to 66.61 mM, and a low detection limit of 0.01 mM or 3.33 mM (S/N = 3), when connected to a potentiostat or a homemade portable biosensor, respectively. The biosensor also exhibited excellent working stability as it retained 82% of its initial performance after 30 days. The analysis of various xylose-containing samples further revealed the merits of our portable xylose biosensor in real-time sensing, including its rapid response, inexpensive instrumentation, and high selectivity, suggesting its great potential in practical applications.