Bioelectrocatalytic activity of reconstituted peroxidase on hemin-terminated PQQ-modified electrodes
Efficient direct electrical communication between an enzyme redox center and an electrode is of great interest from the viewpoints of application to reagentless biosensors and biofuel cells. To immobilize redox enzymes in a favorable orientation for electron transfer at the electrode surface, we rec...
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| Format: | Article |
| Language: | English |
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Elsevier
2022-11-01
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| Series: | Sensors and Actuators Reports |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666053922000534 |
| _version_ | 1811297985104969728 |
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| author | Kikuo Komori Tomoki Aikawa |
| author_facet | Kikuo Komori Tomoki Aikawa |
| author_sort | Kikuo Komori |
| collection | DOAJ |
| description | Efficient direct electrical communication between an enzyme redox center and an electrode is of great interest from the viewpoints of application to reagentless biosensors and biofuel cells. To immobilize redox enzymes in a favorable orientation for electron transfer at the electrode surface, we reconstituted apo-horseradish peroxidase (apo-HRP) onto a hemin cofactor linked to pyrroloquinoline quinone (PQQ) spacer at an indium-tin-oxide (ITO) electrode surface. The bioelectrocatalytic activity of reconstituted HRP for H2O2 reduction was evaluated by means of amperometry at +0.150 V (vs. Ag|AgCl), which value is larger than that for the redox potential of PQQ (ca. –0.140 V). The complex-formation reaction rate of reconstituted HRP with H2O2 enhanced in comparison with that for the hemin linked to PQQ electrode without apo-HRP by a factor of 200. PQQ served as the part of spacer for the insertion of hemin into a certain desired position in apo-HRP. Although native HRP cannot electrochemically communicate with the ITO electrode in general, the quasi-direct electron transfer from the ITO electrode to reconstituted HRP oxidized by H2O2 was also achieved most likely by electron hopping and/or resonant transport without the redox mediation thanks to the π-conjugated PQQ molecule in the spacer. |
| first_indexed | 2024-04-13T06:13:10Z |
| format | Article |
| id | doaj.art-05eb032b69234b1f9595242566548f3d |
| institution | Directory Open Access Journal |
| issn | 2666-0539 |
| language | English |
| last_indexed | 2024-04-13T06:13:10Z |
| publishDate | 2022-11-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Sensors and Actuators Reports |
| spelling | doaj.art-05eb032b69234b1f9595242566548f3d2022-12-22T02:58:56ZengElsevierSensors and Actuators Reports2666-05392022-11-014100126Bioelectrocatalytic activity of reconstituted peroxidase on hemin-terminated PQQ-modified electrodesKikuo Komori0Tomoki Aikawa1Department of Biotechnology and Chemistry, Kindai University, Takaya-Umenobe, Higashi-Hiroshima 739-2116 Japan; Graduate School of System Engineering, Kindai University, Takaya-Umenobe, Higashi-Hiroshima 739-2116 Japan; Corresponding author at: Department of Biotechnology and Chemistry, Kindai University, Takaya-Umenobe, Higashi-Hiroshima 739-2116 Japan.Department of Biotechnology and Chemistry, Kindai University, Takaya-Umenobe, Higashi-Hiroshima 739-2116 JapanEfficient direct electrical communication between an enzyme redox center and an electrode is of great interest from the viewpoints of application to reagentless biosensors and biofuel cells. To immobilize redox enzymes in a favorable orientation for electron transfer at the electrode surface, we reconstituted apo-horseradish peroxidase (apo-HRP) onto a hemin cofactor linked to pyrroloquinoline quinone (PQQ) spacer at an indium-tin-oxide (ITO) electrode surface. The bioelectrocatalytic activity of reconstituted HRP for H2O2 reduction was evaluated by means of amperometry at +0.150 V (vs. Ag|AgCl), which value is larger than that for the redox potential of PQQ (ca. –0.140 V). The complex-formation reaction rate of reconstituted HRP with H2O2 enhanced in comparison with that for the hemin linked to PQQ electrode without apo-HRP by a factor of 200. PQQ served as the part of spacer for the insertion of hemin into a certain desired position in apo-HRP. Although native HRP cannot electrochemically communicate with the ITO electrode in general, the quasi-direct electron transfer from the ITO electrode to reconstituted HRP oxidized by H2O2 was also achieved most likely by electron hopping and/or resonant transport without the redox mediation thanks to the π-conjugated PQQ molecule in the spacer.http://www.sciencedirect.com/science/article/pii/S2666053922000534Enzyme orientationπ-Conjugated moleculesEnzyme activity regulationKinetic analysisHeme proteins |
| spellingShingle | Kikuo Komori Tomoki Aikawa Bioelectrocatalytic activity of reconstituted peroxidase on hemin-terminated PQQ-modified electrodes Sensors and Actuators Reports Enzyme orientation π-Conjugated molecules Enzyme activity regulation Kinetic analysis Heme proteins |
| title | Bioelectrocatalytic activity of reconstituted peroxidase on hemin-terminated PQQ-modified electrodes |
| title_full | Bioelectrocatalytic activity of reconstituted peroxidase on hemin-terminated PQQ-modified electrodes |
| title_fullStr | Bioelectrocatalytic activity of reconstituted peroxidase on hemin-terminated PQQ-modified electrodes |
| title_full_unstemmed | Bioelectrocatalytic activity of reconstituted peroxidase on hemin-terminated PQQ-modified electrodes |
| title_short | Bioelectrocatalytic activity of reconstituted peroxidase on hemin-terminated PQQ-modified electrodes |
| title_sort | bioelectrocatalytic activity of reconstituted peroxidase on hemin terminated pqq modified electrodes |
| topic | Enzyme orientation π-Conjugated molecules Enzyme activity regulation Kinetic analysis Heme proteins |
| url | http://www.sciencedirect.com/science/article/pii/S2666053922000534 |
| work_keys_str_mv | AT kikuokomori bioelectrocatalyticactivityofreconstitutedperoxidaseonheminterminatedpqqmodifiedelectrodes AT tomokiaikawa bioelectrocatalyticactivityofreconstitutedperoxidaseonheminterminatedpqqmodifiedelectrodes |