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Natural enzymes often face limitations related to low stability and high production costs, prompting the development of nanomaterials that mimic enzymatic activity. A copper single-atom nanozyme, designated as Cu-N-C or Cu-SAN, has been developed to replicate the function of natural galactose oxidase. The material exhibits both peroxidase-like and enhanced oxidase-like properties. It displays stereospecific catalytic activity, successfully oxidising D-galactose and primary alcohols while remaining inactive towards L-galactose and other carbohydrates. In the presence of oxygen, the nanozyme catalyses galactose oxidation to yield hydrogen peroxide, which subsequently reacts with a chromogenic substrate to produce a measurable blue colour. The resulting sensor shows a linear response for galactose concentrations between 1 and 60 micromolar, achieving a detection limit of 0.23 micromolar. Theoretical calculations confirm the catalytic mechanisms and explain the selectivity towards D-galactose.
Natural enzymes used in diagnostic testing and food monitoring can be fragile and expensive to produce. Developing synthetic single-atom materials that accurately replicate natural enzyme behaviour offers a more robust alternative. This approach enables reliable, highly specific detection of sugars like D-galactose, which is vital for diagnosing metabolic conditions and verifying product quality in the dairy sector.
This research could enable the production of stable, low-cost diagnostic tests and quality control tools. Potential users include clinical laboratories screening for the metabolic disorder galactosemia, as well as manufacturers testing for galactose levels in dairy and other commercial goods. As the abstract demonstrates analytical performance in laboratory tests, the technology represents early-stage to applied development that requires further real-sample validation before commercial use.
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Due to the low stability and high cost of some natural enzymes, nanozymes have been developed as enzyme-imitating nanomaterials. Single-atom nanozymes are a class of nanozymes with metal centers that mimic the structure of metal-based natural enzymes. Herein, Cu-N-C single-atom nanozyme (SAN) is synthesized with excellent peroxidase- and enhanced oxidase-like activities to mimic the action of natural galactose oxidase. Cu-SAN demonstrates stereospecific activity akin to that of natural galactose oxidase by oxidizing D-galactose and primary alcohol but not L-Galactose or other carbohydrates. The SAN can catalyze the oxidation of galactose in the presence of oxygen, producing hydrogen peroxide as a sub-product. The produced hydrogen peroxide then oxidizes 3,3',5,5'-tetramethylbenzidine catalyzed by the SAN, yielding the typical blue product. The relationship between absorbance and galactose concentration is linear in the 1-60 µm range with a detection limit as low as 0.23 µm. This strategy can be utilized in the diagnosis of galactosemia disorder and detection of galactose in some dairy and other commercial products. DFT calculations clarify the high activity of the Cu sites in the POD-like reaction and explain the selectivity of the Cu-SAN oxidase-like reaction toward D-galactose.
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DOI: 10.1002/smll.202405986
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