Scalable Printing of Prussian Blue Analogue@Au Edge-Rich Microcubes as Flexible Biosensing Microchips Performing Ultrasensitive Sucrose Fermentation Monitoring

Sucrose is one of the most applied carbon sources in the fermentation process, and it directly determines the microbial metabolism with its concentration fluctuation. Meanwhile, sucrose also plays a key role of a protective agent in the production of biological vaccines, especially in the new mRNA vaccines for curing COVID-19. However, rapid and precise detection of sucrose is always desired but unrealized in industrial fermentation and synthetic biology research. In order to address the above issue, we proposed an ultrasensitive biosensor microchip achieving accurate sucrose recognition within only 12 s, relying on the construction of a Prussian blue analogue@Au edge-rich (PBA@AuER) microarchitecture. This special geometric structure was formed through exactly inducing the oriented PBA crystallization toward a certain plane to create more regular and continuous edge features. This composite was further transformed to a screen-printed ink to directly and large-scale fabricate an enzymatic biosensor microchip showing ultrahigh sensitivity, a wide detection range, and a low detection limit to the accurate sucrose recognition. As confirmed in a real alcohol fermentation reaction, the as-prepared microchip enabled us to accurately detect the sucrose and glucose concentrations with outstanding reusability (more than 300 times) during the whole process through proposing a novel analytical strategy for the binary mixture substrate system.