FAD dependent glucose dehydrogenases - Discovery and engineering of representative glucose sensing enzymes.

The history of the development of glucose sensors goes hand-in-hand with the history of the discovery and the engineering of glucose-sensing enzymes. Glucose oxidase (GOx) has been used for glucose sensing since the development of the first electrochemical glucose sensor. The principle utilizing oxygen as the electron acceptor is designated as the first-generation electrochemical enzyme sensors. With increasing demand for hand-held and cost-effective devices for the "self-monitoring of blood glucose (SMBG)", second-generation electrochemical sensor strips employing electron mediators have become the most popular platform. To overcome the inherent drawback of GOx, namely, the use of oxygen as the electron acceptor, various glucose dehydrogenases (GDHs) have been utilized in second-generation principle-based sensors. Among the various enzymes employed in glucose sensors, GDHs harboring FAD as the redox cofactor, FADGDHs, especially those derived from fungi, fFADGDHs, are currently the most popular enzymes in the sensor strips of second-generation SMBG sensors. In addition, the third-generation principle, employing direct electron transfer (DET), is considered the most elegant approach and is ideal for use in electrochemical enzyme sensors. However, glucose oxidoreductases capable of DET are limited. One of the most prominent GDHs capable of DET is a bacteria-derived FADGDH complex (bFADGDH). bFADGDH has three distinct subunits; the FAD harboring the catalytic subunit, the small subunit, and the electron-transfer subunit, which makes bFADGDH capable of DET. In this review, we focused on the two representative glucose sensing enzymes, fFADGDHs and bFADGDHs, by presenting their discovery, sources, and protein and enzyme properties, and the current engineering strategies to improve their potential in sensor applications.