Effect of deglycosylation on the mediated electrocatalytic activity of recombinantly expressed Agaricus meleagris pyranose dehydrogenase wired by osmium redox polymer

Anikó Killyéni; Maria E. Yakovleva; Domhnall Macaodha; Peter Ó Conghaile; Christoph Gonaus; Roberto Ortiz; Dónal Leech; Ionel Catalin Popescu; Clemens K. Peterbauer; Lo Gorton

doi:10.1016/j.electacta.2013.08.069

Effect of deglycosylation on the mediated electrocatalytic activity of recombinantly expressed Agaricus meleagris pyranose dehydrogenase wired by osmium redox polymer

Anikó Killyéni, Maria E. Yakovleva, Domhnall Macaodha, Peter Ó Conghaile, Christoph Gonaus, Roberto Ortiz, Dónal Leech, Ionel Catalin Popescu, Clemens K. Peterbauer, Lo Gorton

Cellular & Molecular Medicine

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12 Citations (Scopus)

Abstract

The effect of deglycosylation of pyranose dehydrogenase (PDH) obtained from Agaricus meleagris (Am) and recombinantly expressed in Pichia pastoris on its electrocatalytic activity was investigated. Glycosylated (gAmPDH) and deglycosylated PDH (dgAmPDH) were immobilised on spectrographic graphite (G) simultaneously with an osmium redox polymer (Os-RP) using poly(ethylene glycol)(400) diglycidyl ether (PEGDGE) as cross-linking agent. The amperometric response to glucose, recorded at G/(Os-RP)-gAmPDH and G/(Os-RP)-dgAmPDH bioelectrodes, was optimised under flow injection conditions concerning the applied potential, enzyme loading, working pH and flow rate. The G/(Os-RP)-dgAmPDH bioelectrode is characterised by better kinetic and electroanalytical parameters compared with the G/(Os-RP)-gAmPDH bioelectrode: (i) a higher value of the maximum catalytic current density, j_max = (146.6 ± 2.6) μA cm^-2 vs. j_max = (80.9 ± 1.9) μA cm^-2; (ii) a lower value of the apparent Michaelis-Menten constant, KMapp=2.4±0.1.

Original language	English
Pages (from-to)	61-67
Number of pages	7
Journal	Electrochimica Acta
Volume	126
DOIs	https://doi.org/10.1016/j.electacta.2013.08.069
Publication status	Published - 20 Apr 2014

Keywords

Enzyme deglycosylation
Mediated electron transfer
Osmium redox polymer
Pyranose dehydrogenase
Substrate selectivity

Authors (Note for portal: view the doc link for the full list of authors)

Authors
Killyeni, A,Yakovleva, ME,MacAodha, D,Conghaile, PO,Gonaus, C,Ortiz, R,Leech, D,Popescu, IC,Peterbauer, CK,Gorton, L

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10.1016/j.electacta.2013.08.069

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Killyéni, A., Yakovleva, M. E., Macaodha, D., Conghaile, P. Ó., Gonaus, C., Ortiz, R., Leech, D., Popescu, I. C., Peterbauer, C. K., & Gorton, L. (2014). Effect of deglycosylation on the mediated electrocatalytic activity of recombinantly expressed Agaricus meleagris pyranose dehydrogenase wired by osmium redox polymer. Electrochimica Acta, 126, 61-67. https://doi.org/10.1016/j.electacta.2013.08.069

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title = "Effect of deglycosylation on the mediated electrocatalytic activity of recombinantly expressed Agaricus meleagris pyranose dehydrogenase wired by osmium redox polymer",

abstract = "The effect of deglycosylation of pyranose dehydrogenase (PDH) obtained from Agaricus meleagris (Am) and recombinantly expressed in Pichia pastoris on its electrocatalytic activity was investigated. Glycosylated (gAmPDH) and deglycosylated PDH (dgAmPDH) were immobilised on spectrographic graphite (G) simultaneously with an osmium redox polymer (Os-RP) using poly(ethylene glycol)(400) diglycidyl ether (PEGDGE) as cross-linking agent. The amperometric response to glucose, recorded at G/(Os-RP)-gAmPDH and G/(Os-RP)-dgAmPDH bioelectrodes, was optimised under flow injection conditions concerning the applied potential, enzyme loading, working pH and flow rate. The G/(Os-RP)-dgAmPDH bioelectrode is characterised by better kinetic and electroanalytical parameters compared with the G/(Os-RP)-gAmPDH bioelectrode: (i) a higher value of the maximum catalytic current density, jmax = (146.6 ± 2.6) μA cm-2 vs. jmax = (80.9 ± 1.9) μA cm-2; (ii) a lower value of the apparent Michaelis-Menten constant, KMapp=2.4±0.1.",

keywords = "Enzyme deglycosylation, Mediated electron transfer, Osmium redox polymer, Pyranose dehydrogenase, Substrate selectivity",

author = "Anik{\'o} Killy{\'e}ni and Yakovleva, \{Maria E.\} and Domhnall Macaodha and Conghaile, \{Peter {\'O}\} and Christoph Gonaus and Roberto Ortiz and D{\'o}nal Leech and Popescu, \{Ionel Catalin\} and Peterbauer, \{Clemens K.\} and Lo Gorton",

year = "2014",

month = apr,

day = "20",

doi = "10.1016/j.electacta.2013.08.069",

language = "English",

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journal = "Electrochimica Acta",

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Killyéni, A, Yakovleva, ME, Macaodha, D, Conghaile, PÓ, Gonaus, C, Ortiz, R, Leech, D, Popescu, IC, Peterbauer, CK & Gorton, L 2014, 'Effect of deglycosylation on the mediated electrocatalytic activity of recombinantly expressed Agaricus meleagris pyranose dehydrogenase wired by osmium redox polymer', Electrochimica Acta, vol. 126, pp. 61-67. https://doi.org/10.1016/j.electacta.2013.08.069

Effect of deglycosylation on the mediated electrocatalytic activity of recombinantly expressed Agaricus meleagris pyranose dehydrogenase wired by osmium redox polymer. / Killyéni, Anikó; Yakovleva, Maria E.; Macaodha, Domhnall et al.
In: Electrochimica Acta, Vol. 126, 20.04.2014, p. 61-67.

Research output: Contribution to a Journal (Peer & Non Peer) › Article › peer-review

TY - JOUR

T1 - Effect of deglycosylation on the mediated electrocatalytic activity of recombinantly expressed Agaricus meleagris pyranose dehydrogenase wired by osmium redox polymer

AU - Killyéni, Anikó

AU - Yakovleva, Maria E.

AU - Macaodha, Domhnall

AU - Conghaile, Peter Ó

AU - Gonaus, Christoph

AU - Ortiz, Roberto

AU - Leech, Dónal

AU - Popescu, Ionel Catalin

AU - Peterbauer, Clemens K.

AU - Gorton, Lo

PY - 2014/4/20

Y1 - 2014/4/20

N2 - The effect of deglycosylation of pyranose dehydrogenase (PDH) obtained from Agaricus meleagris (Am) and recombinantly expressed in Pichia pastoris on its electrocatalytic activity was investigated. Glycosylated (gAmPDH) and deglycosylated PDH (dgAmPDH) were immobilised on spectrographic graphite (G) simultaneously with an osmium redox polymer (Os-RP) using poly(ethylene glycol)(400) diglycidyl ether (PEGDGE) as cross-linking agent. The amperometric response to glucose, recorded at G/(Os-RP)-gAmPDH and G/(Os-RP)-dgAmPDH bioelectrodes, was optimised under flow injection conditions concerning the applied potential, enzyme loading, working pH and flow rate. The G/(Os-RP)-dgAmPDH bioelectrode is characterised by better kinetic and electroanalytical parameters compared with the G/(Os-RP)-gAmPDH bioelectrode: (i) a higher value of the maximum catalytic current density, jmax = (146.6 ± 2.6) μA cm-2 vs. jmax = (80.9 ± 1.9) μA cm-2; (ii) a lower value of the apparent Michaelis-Menten constant, KMapp=2.4±0.1.

AB - The effect of deglycosylation of pyranose dehydrogenase (PDH) obtained from Agaricus meleagris (Am) and recombinantly expressed in Pichia pastoris on its electrocatalytic activity was investigated. Glycosylated (gAmPDH) and deglycosylated PDH (dgAmPDH) were immobilised on spectrographic graphite (G) simultaneously with an osmium redox polymer (Os-RP) using poly(ethylene glycol)(400) diglycidyl ether (PEGDGE) as cross-linking agent. The amperometric response to glucose, recorded at G/(Os-RP)-gAmPDH and G/(Os-RP)-dgAmPDH bioelectrodes, was optimised under flow injection conditions concerning the applied potential, enzyme loading, working pH and flow rate. The G/(Os-RP)-dgAmPDH bioelectrode is characterised by better kinetic and electroanalytical parameters compared with the G/(Os-RP)-gAmPDH bioelectrode: (i) a higher value of the maximum catalytic current density, jmax = (146.6 ± 2.6) μA cm-2 vs. jmax = (80.9 ± 1.9) μA cm-2; (ii) a lower value of the apparent Michaelis-Menten constant, KMapp=2.4±0.1.

KW - Enzyme deglycosylation

KW - Mediated electron transfer

KW - Osmium redox polymer

KW - Pyranose dehydrogenase

KW - Substrate selectivity

UR - https://www.scopus.com/pages/publications/84897995460

U2 - 10.1016/j.electacta.2013.08.069

DO - 10.1016/j.electacta.2013.08.069

M3 - Article

SN - 0013-4686

VL - 126

SP - 61

EP - 67

JO - Electrochimica Acta

JF - Electrochimica Acta

ER -

Effect of deglycosylation on the mediated electrocatalytic activity of recombinantly expressed Agaricus meleagris pyranose dehydrogenase wired by osmium redox polymer

Abstract

Keywords

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