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A highly selective biosensor with nanomolar sensitivity based on cytokinin dehydrogenase
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Tian, Faming, Greplová, Marta, Frébort, Ivo, Dale, Nicholas and Napier, R. (Richard) (2014) A highly selective biosensor with nanomolar sensitivity based on cytokinin dehydrogenase. PLoS One, Volume 9 (Number 3). Article number e90877. doi:10.1371/journal.pone.0090877
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Official URL: http://dx.doi.org/10.1371/journal.pone.0090877
Abstract
We have developed a N6-dimethylallyladenine (cytokinin) dehydrogenase-based microbiosensor for real-time determination of the family of hormones known as cytokinins. Cytokinin dehydrogenase from Zea mays (ZmCKX1) was immobilised concurrently with electrodeposition of a silica gel film on the surface of a Pt microelectrode, which was further functionalized by free electron mediator 2,6-dichlorophenolindophenol (DCPIP) in supporting electrolyte to give a bioactive film capable of selective oxidative cleavage of the N6- side chain of cytokinins. The rapid electron shuffling between freely diffusible DCPIP and the FAD redox group in ZmCKX1 endowed the microbiosensor with a fast response time of less than 10 s. The immobilised ZmCKX1 retained a high affinity for its preferred substrate N6-(Δ2-isopentenyl) adenine (iP), and gave the miniaturized biosensor a large linear dynamic range from 10 nM to 10 µM, a detection limit of 3.9 nM and a high sensitivity to iP of 603.3 µAmM−1cm−2 (n = 4, R2 = 0.9999). Excellent selectivity was displayed for several other aliphatic cytokinins and their ribosides, including N6-(Δ2-isopentenyl) adenine, N6-(Δ2-isopentenyl) adenosine, cis-zeatin, trans-zeatin and trans-zeatin riboside. Aromatic cytokinins and metabolites such as cytokinin glucosides were generally poor substrates. The microbiosensors exhibited excellent stability in terms of pH and long-term storage and have been used successfully to determine low nanomolar cytokinin concentrations in tomato xylem sap exudates.
| Item Type: | Journal Article | ||||||||
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| Subjects: | Q Science > QK Botany | ||||||||
| Divisions: | Faculty of Science > Life Sciences (2010- ) | ||||||||
| Library of Congress Subject Headings (LCSH): | Cytokinins, Biosensors | ||||||||
| Journal or Publication Title: | PLoS One | ||||||||
| Publisher: | Public Library of Science | ||||||||
| ISSN: | 1932-6203 | ||||||||
| Official Date: | 4 March 2014 | ||||||||
| Dates: |
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| Volume: | Volume 9 | ||||||||
| Number: | Number 3 | ||||||||
| Article Number: | Article number e90877 | ||||||||
| DOI: | 10.1371/journal.pone.0090877 | ||||||||
| Status: | Peer Reviewed | ||||||||
| Publication Status: | Published | ||||||||
| Access rights to Published version: | Open Access | ||||||||
| Funder: | Biotechnology and Biological Sciences Research Council (Great Britain) (BBSRC), European Social Fund (ESF) | ||||||||
| Grant number: | BB/I023933/1 (BBSRC), CZ.1.07/2.3.00/20.0165 (ESF) |
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