Employment of 1-Methoxy-5-Ethyl Phenazinium Ethyl Sulfate as a Stable Electron Mediator in Flavin Oxidoreductases-Based Sensors

Sensors (Basel). 2020 May 15;20(10):2825. doi: 10.3390/s20102825.

Abstract

In this paper, a novel electron mediator, 1-methoxy-5-ethyl phenazinium ethyl sulfate (mPES), was introduced as a versatile mediator for disposable enzyme sensor strips, employing representative flavin oxidoreductases, lactate oxidase (LOx), glucose dehydrogenase (GDH), and fructosyl peptide oxidase (FPOx). A disposable lactate enzyme sensor with oxygen insensitive Aerococcus viridans-derived engineered LOx (AvLOx), with A96L mutant as the enzyme, was constructed. The constructed lactate sensor exhibited a high sensitivity (0.73 ± 0.12 μA/mM) and wide linear range (0-50 mM lactate), showings that mPES functions as an effective mediator for AvLOx. Employing mPES as mediator allowed this amperometric lactate sensor to be operated at a relatively low potential of +0.2 V to 0 V vs. Ag/AgCl, thus avoiding interference from uric acid and acetaminophen. The lactate sensors were adequately stable for at least 48 days of storage at 25 °C. These results indicated that mPES can be replaced with 1-methoxy-5-methyl phenazinium methyl sulfate (mPMS), which we previously reported as the best mediator for AvLOx-based lactate sensors. Furthermore, this study revealed that mPES can be used as an effective electron mediator for the enzyme sensors employing representative flavin oxidoreductases, GDH-based glucose sensors, and FPOx-based hemoglobin A1c (HbA1c) sensors.

Keywords: 1-methoxy-5-ethyl phenazinium ethyl sulfate; biomedical engineering; disposable enzyme sensor; electrochemical enzyme sensor; fructosyl peptide oxidase; glucose dehydrogenase; lactate oxidase.

MeSH terms

  • Aerococcus / enzymology*
  • Amino Acid Oxidoreductases / chemistry*
  • Biosensing Techniques*
  • Electrons*
  • Glucose Dehydrogenases / chemistry*
  • Mixed Function Oxygenases / chemistry*
  • Sulfuric Acid Esters / chemistry

Substances

  • Sulfuric Acid Esters
  • Mixed Function Oxygenases
  • Glucose Dehydrogenases
  • lactate 2-monooxygenase
  • Amino Acid Oxidoreductases
  • fructosyl-peptide oxidase
  • diethyl sulfate