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Synergistic Bioinorganic Interfaces: Laccase-Like Nanozymes Coupled with Natural Laccase as Stable and High-Performance Cathodic Catalysts in Enzymatic Biofuel Cells

  • Mykhailo Gonchar 1,3,*,   
  • Galina Gayda 1,   
  • Halyna Klepach 3,   
  • Oksana Zakalska 1,   
  • Nataliya Stasyuka 1,2,   
  • Demkiv Olha 1,2,*

Received: 04 Nov 2025 | Revised: 18 Nov 2025 | Accepted: 24 Nov 2025 | Published: 05 Dec 2025

Abstract

The limited kinetics of the oxygen reduction reaction (ORR) at the cathode remains a key bottleneck in improving the power output of enzymatic biofuel cells (EBFCs). In this study, we present a bioinorganic hybridization that integrates natural laccase (Lac) with laccase-like nanozymes (LacNZs) based on multimetallic composites to improve charge-transfer efficiency. The designed LacNZs structurally and functionally emulate the multicopper catalytic core of laccase, enabling accelerated electron transport and efficient ORR through a synergistic bioinorganic electron-transfer mechanism. EBFCs were fabricated by coupling an AOx/CNT-based bioanode with the constructed enzyme- and nanozyme-based cathodes containing either Lac, LacNZ, or laccase composites Lac/LacNZ, and all systems were evaluated under identical conditions. Incorporation of LacNZs into the cathodic layer resulted in a substantial enhancement of electrocatalytic activity and overall cell performance. The nCuCeAuZIF/Lac/GCE and nCoCuCeZIF/Lac/GCE hybrid cathodes demonstrated power densities of 3.4 μW·cm−2, OCV = 0.610 V, and 2.2 μW·cm−2, OCV = 0.550 V, respectively—nearly one order of magnitude higher than that of the enzyme Lac/CNT/GCE electrode. This improvement arises from the synergistic redox interplay among the multimetallic centers (Co, Cu, Ce, and Au) within the LacNZs, which effectively reproduce the T1–T3 copper sites of native laccase, thereby facilitating a direct four-electron reduction of O2 to H2O, enhancing electron mobility, and stabilizing the enzyme’s conformation at the bioinorganic interface. The hybrid cathode also exhibited outstanding stability: retaining 75% of initial activity after 10 days (vs. 30% for Lac/GCE) and showing a 7.5% increase in sensitivity under 0.25 M NaCl (vs. 41.7% loss for Lac/GCE). Notably, EBFCs with only-LacNZs-based cathodes (nCuCeAu/GCE and nCoCuCeZIF/GCE) retained significant ORR activity (up to 1.2 μW·cm−2 and 0.53 μW·cm−2), highlighting their potential as enzyme-free, durable, and cost-effective catalysts. Nanozymes as catalytic elements of EBFCs can be regarded as an alternative to direct electron transfer technology, which is still complicated for proteins enzymes.

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Gonchar, M.; Gayda, G.; Klepach, H.; Zakalska, O.; Stasyuka, N.; Olha, D. Synergistic Bioinorganic Interfaces: Laccase-Like Nanozymes Coupled with Natural Laccase as Stable and High-Performance Cathodic Catalysts in Enzymatic Biofuel Cells. Bioelectrochemistry and Biosensors 2025, 1 (1), 3.
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