Rapid determination of sucrose and glucose in microbial fermentation and fruit juice samples using engineered multi-enzyme biosensing microchip

Amperometric biosensor utilizing prussian blue (PB) nanocubes microchip for rapid and accurate determination of sucrose and glucose contents in the fermentation broth and juice samples is described. Using a chitosan embedding immobilization strategy, this multi-enzyme system consisting of engineered invertase/mutarotase/glucose oxidase was co-immobilized on a PB nanocubes screen-printed microchip to construct a high-performance and portable sucrose biosensor. Under work potential of −0.05 V, the optimized biosensor showed a fast response time of 20 s and wide linear range from 0.5 to 4.5 mM (R² = 0.998) with a detection limit of 0.02 mM sucrose. Besides, the biosensor showed a strong anti-interference and operational stability with a low RSD (<2%) and 101.03% of average recovery in real E. coli fermentation samples. Finally, the biosensor was implemented together with a glucose amperometric biosensor to perform the simultaneous quantification of sucrose and glucose in commercial fruit juice. These essential characteristics indicated the proposed sucrose biosensor has potential for application in fermentation industry and beverage products.