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Amperometric Flow Injection Analysis of Glucose and Galactose Based on Engineered Pyranose 2-Oxidases and Osmium Polymers for Biosensor Applications

Kurbanoglu, Sevinc; Zafar, Muhammed Nadeem; Tasca, Federico LU ; Aslam, Iqra; Spadiut, Oliver; Leech, Dónal; Haltrich, Dietmar and Gorton, Lo LU (2018) In Electroanalysis 30(7). p.1496-1504
Abstract

In the present study, wild type and three mutants of pyranose 2-oxidase (PyOx-WT, PyOx-MT1, PyOx-MT2, PyOx-MT3), which showed improved properties for D-galactose oxidation, were investigated for their oxidising ability when immobilised on graphite electrodes. Four different flexible Os polymers with formal potentials ranging between −0.140 and 0.270 V vs. Ag|AgCl0.1 M KCl were applied together with the various forms of PyOx to wire graphite electrodes using polyethylene glycol diglycidyl ether as crosslinking reagent. The pH profiles for the electrodes modified with wild type and all PyOx mutants in combination with the Os polymers were investigated with both glucose and galactose, respectively, since the PyOx variants showed... (More)

In the present study, wild type and three mutants of pyranose 2-oxidase (PyOx-WT, PyOx-MT1, PyOx-MT2, PyOx-MT3), which showed improved properties for D-galactose oxidation, were investigated for their oxidising ability when immobilised on graphite electrodes. Four different flexible Os polymers with formal potentials ranging between −0.140 and 0.270 V vs. Ag|AgCl0.1 M KCl were applied together with the various forms of PyOx to wire graphite electrodes using polyethylene glycol diglycidyl ether as crosslinking reagent. The pH profiles for the electrodes modified with wild type and all PyOx mutants in combination with the Os polymers were investigated with both glucose and galactose, respectively, since the PyOx variants showed an improved catalytic activity for galactose. All modified electrodes showed highest response in the pH range between 8.5–10 and KM, Imax values for both glucose and galactose were determined. To prove the catalytic activity, the biosensors were also characterized with cyclic voltammetry. A protein amount 0.26 U was found optimum for PyOx-WT, 0.36 U for PyOx-MT1, 0.41 U for PyOx-MT2 and 0.28 U for PyOx-MT3 and the analytical characterization of the enzyme electrodes was performed for glucose and galactose under optimized conditions.

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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
biofuel cell anode, Biosensor, galactose, glucose, osmium redox polymers, pyranose oxidase
in
Electroanalysis
volume
30
issue
7
pages
9 pages
publisher
John Wiley & Sons
external identifiers
  • scopus:85049859725
ISSN
1040-0397
DOI
10.1002/elan.201800096
language
English
LU publication?
yes
id
a8d7a38f-ff37-4fb3-ad44-c370e9604b77
date added to LUP
2018-07-30 15:02:38
date last changed
2019-02-20 11:23:16
@article{a8d7a38f-ff37-4fb3-ad44-c370e9604b77,
  abstract     = {<p>In the present study, wild type and three mutants of pyranose 2-oxidase (PyOx-WT, PyOx-MT1, PyOx-MT2, PyOx-MT3), which showed improved properties for D-galactose oxidation, were investigated for their oxidising ability when immobilised on graphite electrodes. Four different flexible Os polymers with formal potentials ranging between −0.140 and 0.270 V vs. Ag|AgCl<sub>0.1 M KCl</sub> were applied together with the various forms of PyOx to wire graphite electrodes using polyethylene glycol diglycidyl ether as crosslinking reagent. The pH profiles for the electrodes modified with wild type and all PyOx mutants in combination with the Os polymers were investigated with both glucose and galactose, respectively, since the PyOx variants showed an improved catalytic activity for galactose. All modified electrodes showed highest response in the pH range between 8.5–10 and K<sub>M</sub>, I<sub>max</sub> values for both glucose and galactose were determined. To prove the catalytic activity, the biosensors were also characterized with cyclic voltammetry. A protein amount 0.26 U was found optimum for PyOx-WT, 0.36 U for PyOx-MT1, 0.41 U for PyOx-MT2 and 0.28 U for PyOx-MT3 and the analytical characterization of the enzyme electrodes was performed for glucose and galactose under optimized conditions.</p>},
  author       = {Kurbanoglu, Sevinc and Zafar, Muhammed Nadeem and Tasca, Federico and Aslam, Iqra and Spadiut, Oliver and Leech, Dónal and Haltrich, Dietmar and Gorton, Lo},
  issn         = {1040-0397},
  keyword      = {biofuel cell anode,Biosensor,galactose,glucose,osmium redox polymers,pyranose oxidase},
  language     = {eng},
  month        = {07},
  number       = {7},
  pages        = {1496--1504},
  publisher    = {John Wiley & Sons},
  series       = {Electroanalysis},
  title        = {Amperometric Flow Injection Analysis of Glucose and Galactose Based on Engineered Pyranose 2-Oxidases and Osmium Polymers for Biosensor Applications},
  url          = {http://dx.doi.org/10.1002/elan.201800096},
  volume       = {30},
  year         = {2018},
}