REVIEW: Application of D-Amino Acid Oxidase (DAAO) in Bioanalytics

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Denis L. Atroshenko^1,2,3,a*, Svyatoslav S. Savin¹, Tatyana S. Oretskaya⁴, and Vladimir I. Tishkov¹

¹Department of Chemistry, Lomonosov Moscow State University, 119991 Moscow, Russia

²Laboratory of Molecular Engineering, Federal Research Center “Fundamental Basis of Biotechnology”, Russian Academy of Sciences, 119071 Moscow, Russia

³Institute of Medicine, RUDN University, 117198 Moscow, Russia

⁴Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119991 Moscow, Russia

^* To whom correspondence should be addressed.

Received: October 7, 2025; Revised: October 23, 2025; Accepted: October 30, 2025

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D-Amino acid oxidase (DAAO, EC 1.4.3.3) is a FAD-dependent enzyme that catalyzes the oxidative deamination of D-amino acids to produce the corresponding α-keto acids, hydrogen peroxide, and ammonium ion. High stereoselectivity toward D-enantiomers and favorable kinetic parameters make DAAO a convenient biocatalytic element for analytical applications. This review systematizes the main areas of DAAO use in bioanalysis, including clinical diagnostics, monitoring of food and biotechnological processes, and environmental surveillance. Sensor platforms and detection modes are discussed, including colorimetry, fluorimetry, chemiluminescence, electrochemistry, photoelectrochemistry, and oxygen-based detection methods. The review also addresses factors determining analytical suitability, strategies to broaden selectivity, as well as engineering approaches and structure-guided discovery of new DAAOs. Current limitations are highlighted and future prospects are outlined, such as improving enzyme stability, scaling up portable devices, and integrating biosensing with digital analytics and machine-learning algorithms.

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KEY WORDS: D-amino acids, D-amino acid oxidase (DAAO), bioanalytics, biosensors, clinical diagnostics

DOI: 10.1134/S0006297925604393

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