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We report the single-step fabrication of advanced electrochemical biosensors upon drop-casting of "artificial peroxidase", that is, oxidase-containing mixtures. Compared to bienzyme biosensors, we substitute peroxidase with more active and stable nanozymes based on catalytically synthesized Prussian Blue. The electroactivity of nanozymes in the sensing membrane was facilitated with carbon black nanoparticles, which resulted in record sensitivity of the H 2 O 2 transducer exceeding 1.1 A M À 1 • cm À 2. Both glucose and lactate oxidases were co-immobilized with nanozymes in alkoxysilane or perfluorosulfonated ionomer membranes. The proposed drop-casting approach results in biosensors that are advantageous over those conventionally produced upon layer-by-layer immobilization in terms of i) almost an order of magnitude increased sensitivity, ii) unsurpassed biosensor-to-transducer sensitivity ratio reaching the value of 0.3, and iii) three times extended operational lifetime.

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