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Characterizing Mediated Extracellular Electron Transfer in Lactic Acid Bacteria with a Three-Electrode, Two-Chamber Bioelectrochemical System

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Optimization and characterization of a lactate-oxidase electrode.

Ke Shi¹, Selvarajan Varshini¹, Keerthi Booshan Manikandan¹

¹Department of Chemical Engineering and RIGET, Gyeongsang National University 501 Jinju-daero Jinju Gyeongnam 660-701 Republic of Korea cj_kim@gnu.ac.kr.

|November 6, 2025

Summary

Researchers optimized lactate oxidase electrodes for biosensors by adjusting enzyme loading and layering. This enhanced electrode performance, crucial for accurate lactate detection in healthcare and sports applications.

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Area of Science:

Electrochemistry
Biosensor Technology
Biomaterials

Background:

Electrochemical lactate sensors are vital for analyzing biological fluids in healthcare and sports.
The performance of these sensors heavily relies on the lactate oxidase (LOx)-based electrode.

Purpose of the Study:

To enhance the performance of lactate oxidase electrodes.
To optimize the loading and layering of lactate oxidase (LOx) and poly(ethylene glycol) diglycidyl ether (PEGDGE) on carbon paper.

Main Methods:

Utilized the Box-Behnken design for optimization.
Characterized the electrode surface using FE-SEM, FTIR, and impedance analyses.
Evaluated performance through electrochemical and biochemical analyses.

Main Results:

The optimized electrode (4 layers LOx, 184 μg PEGDGE) achieved an oxidation current of 1840 ± 60 μA.
Demonstrated high enzyme activity, a maximum current at 50 mM lactate, and an apparent K^app_m of 11.4 mM.
Exhibited high stability and robust enzyme binding for repeated use.

Conclusions:

Optimized LOx/PEGDGE loading and layering significantly enhance electrode performance.
The developed electrode is suitable for creating more effective and dependable lactate biosensors.
This work contributes to advancements in electrochemical biosensing for biological fluid analysis.