Development of closed bipolar electrode based L-lactate sensor employing quasi-direct electron transfer type enzyme with cyclic voltammetry

Abstract

Herein, we present a proof-of-concept of an enzyme sensor combining closed bipolar electrode system with quasi-direct electron transfer (DET) type enzyme. The closed bipolar electrode system was tested using cyclic voltammetry, with L-lactate as a model substrate. L-Lactate was detected through measurement of the change in junction potential across the bipolar electrode. This change in junction potential was caused by reduction of amino reactive phenazine ethosulfate conjugated to Aerococcus vilidans derived engineered L-lactate oxidase (AvLOx) which shows a quasi-DET signal. Using the closed bipolar electrode system allowed simultaneous measuring using cyclic voltammetry and open circuit potential (OCP) and achieved a limit of detection of 400 μM and 76.2 μM lactate respectively. The sensor was then demonstrated to perform with equivalent sensitivity using OCP across varying surface areas. To the best of our knowledge this is the first time a closed bipolar electrode system has been used with an enzyme which is capable of quasi-direct or direct electron transfer. This work can be expanded further to other enzymes capable of directly altering the junction potential of an electrode surface.