The detection of small molecules has attracted enormous interest in various fields, including the chemical, biological, and healthcare fields. In order to achieve such detection with high accuracy, up to now, various types of biosensors have been developed. Among those biosensors, enzymatic biosensors have shown excellent sensing performances via their highly specific enzymatic reactions with small chemical molecules. As techniques used to implement the sensing function of such enzymatic biosensors, electrochemical and fluorescence techniques have been mostly used for the detection of small molecules because of their advantages. In addition, through the incorporation of nanotechnologies, the detection property of each technique-based enzymatic nanobiosensors can be improved to measure harmful or important small molecules accurately. This review provides interdisciplinary information related to developing enzymatic nanobiosensors for small molecule detection, such as widely used enzymes, target small molecules, and electrochemical/fluorescence techniques. We expect that this review will provide a broad perspective and well-organized roadmap to develop novel electrochemical and fluorescent enzymatic nanobiosensors.
