Direct electrochemistry and biolelectrocatalysis of H2O2 reduction of recombinant tobacco peroxidase on graphite. Effect of peroxidase single-point mutation on Ca2+-modulated catalytic activity
(2006) In Journal of Electroanalytical Chemistry 588(1). p.112-121- Abstract
- Direct electron transfer (DET) reactions and bio(electro)catalytic reduction of H2O2 catalysed by native and recombinant forms of tobacco peroxidase (nTOP and rTOP) were studied in homogeneous-phase catalysis and when TOPs were adsorbed on graphite electrodes. Non-glycosylated wild type and Glu141 -> Phe mutant forms of rTOP were produced using an Escherichia coli expression system. Mutation was introduced to explore the mechanisms for modulation of the catalytic activity of TOP by Ca2+ ions. At the pH optimum of 5.0, direct electrochemical Fe3+/2+ transformation of the peroxidase heme was characterised by potentials of -208 mV (nTOP) and -239 mV vs. Ag vertical bar AgCl (rTOP), and 0.9 +/- 0.1 and 1.1 +/- 0.4 pmoles of adsorbed nTOP... (More)
- Direct electron transfer (DET) reactions and bio(electro)catalytic reduction of H2O2 catalysed by native and recombinant forms of tobacco peroxidase (nTOP and rTOP) were studied in homogeneous-phase catalysis and when TOPs were adsorbed on graphite electrodes. Non-glycosylated wild type and Glu141 -> Phe mutant forms of rTOP were produced using an Escherichia coli expression system. Mutation was introduced to explore the mechanisms for modulation of the catalytic activity of TOP by Ca2+ ions. At the pH optimum of 5.0, direct electrochemical Fe3+/2+ transformation of the peroxidase heme was characterised by potentials of -208 mV (nTOP) and -239 mV vs. Ag vertical bar AgCl (rTOP), and 0.9 +/- 0.1 and 1.1 +/- 0.4 pmoles of adsorbed nTOP and rTOP, correspondingly, were in DET contact with graphite. Kinetic analysis of amperometric (at +50 mV) data on H2O2 reduction at TOP-modified electrodes, placed in a wall-jet flow-through electrochemical cell, yielded 82% (nTOP) and 88% (rTOP) of adsorbed TOP molecules active in the DET reaction. The efficiency of DET (and bioelectrocatalysis) increased 3.5-fold when changing from glycosylated nTOP to rTOP. The Glu141 Phe mutation in the heme-binding pocket of rTOP enabled to achieve a Ca2+-tolerance of TOP in the reaction with H2O2, which is characteristic of other plant peroxidases, and to a large extent in heterogeneous DET and reaction with a second substrate catechol. The results promote further applications of TOP for biosensor- and solid-phase biocatalysts development. (c) 2005 Elsevier B.V. All rights reserved. (Less)
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https://lup.lub.lu.se/record/417783
- author
- Castillo, John LU ; Ferapontova, Elena LU ; Hushpulian, D ; Tasca, Federico LU ; Tishkov, V ; Chubar, T ; Gazaryan, I and Gorton, Lo LU
- organization
- publishing date
- 2006
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- heterogeneous direct electron transfer, graphite electrodes, refolding, single-point mutation, peroxidase, recombinant tobacco, enzymatic catalysis, bioelectrocatalysis
- in
- Journal of Electroanalytical Chemistry
- volume
- 588
- issue
- 1
- pages
- 112 - 121
- publisher
- Elsevier
- external identifiers
-
- wos:000235264000013
- scopus:31344465504
- ISSN
- 1572-6657
- DOI
- 10.1016/j.jelechem.2005.12.010
- language
- English
- LU publication?
- yes
- additional info
- The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Analytical Chemistry (S/LTH) (011001004)
- id
- 33288013-9dab-460e-9f91-bba252bbd546 (old id 417783)
- date added to LUP
- 2016-04-01 16:43:20
- date last changed
- 2022-01-28 21:39:31
@article{33288013-9dab-460e-9f91-bba252bbd546,
abstract = {{Direct electron transfer (DET) reactions and bio(electro)catalytic reduction of H2O2 catalysed by native and recombinant forms of tobacco peroxidase (nTOP and rTOP) were studied in homogeneous-phase catalysis and when TOPs were adsorbed on graphite electrodes. Non-glycosylated wild type and Glu141 -> Phe mutant forms of rTOP were produced using an Escherichia coli expression system. Mutation was introduced to explore the mechanisms for modulation of the catalytic activity of TOP by Ca2+ ions. At the pH optimum of 5.0, direct electrochemical Fe3+/2+ transformation of the peroxidase heme was characterised by potentials of -208 mV (nTOP) and -239 mV vs. Ag vertical bar AgCl (rTOP), and 0.9 +/- 0.1 and 1.1 +/- 0.4 pmoles of adsorbed nTOP and rTOP, correspondingly, were in DET contact with graphite. Kinetic analysis of amperometric (at +50 mV) data on H2O2 reduction at TOP-modified electrodes, placed in a wall-jet flow-through electrochemical cell, yielded 82% (nTOP) and 88% (rTOP) of adsorbed TOP molecules active in the DET reaction. The efficiency of DET (and bioelectrocatalysis) increased 3.5-fold when changing from glycosylated nTOP to rTOP. The Glu141 Phe mutation in the heme-binding pocket of rTOP enabled to achieve a Ca2+-tolerance of TOP in the reaction with H2O2, which is characteristic of other plant peroxidases, and to a large extent in heterogeneous DET and reaction with a second substrate catechol. The results promote further applications of TOP for biosensor- and solid-phase biocatalysts development. (c) 2005 Elsevier B.V. All rights reserved.}},
author = {{Castillo, John and Ferapontova, Elena and Hushpulian, D and Tasca, Federico and Tishkov, V and Chubar, T and Gazaryan, I and Gorton, Lo}},
issn = {{1572-6657}},
keywords = {{heterogeneous direct electron transfer; graphite electrodes; refolding; single-point mutation; peroxidase; recombinant tobacco; enzymatic catalysis; bioelectrocatalysis}},
language = {{eng}},
number = {{1}},
pages = {{112--121}},
publisher = {{Elsevier}},
series = {{Journal of Electroanalytical Chemistry}},
title = {{Direct electrochemistry and biolelectrocatalysis of H2O2 reduction of recombinant tobacco peroxidase on graphite. Effect of peroxidase single-point mutation on Ca2+-modulated catalytic activity}},
url = {{http://dx.doi.org/10.1016/j.jelechem.2005.12.010}},
doi = {{10.1016/j.jelechem.2005.12.010}},
volume = {{588}},
year = {{2006}},
}