A highly selective biosensor with nanomolar sensitivity based on cytokinin dehydrogenase.

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...

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Autores principales:	Faming Tian, Marta Greplová, Ivo Frébort, Nicholas Dale, Richard Napier
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/706fdac6606d4fa8a841e3921766d00f
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spelling	oai:doaj.org-article:706fdac6606d4fa8a841e3921766d00f2021-11-18T08:29:46ZA highly selective biosensor with nanomolar sensitivity based on cytokinin dehydrogenase.1932-620310.1371/journal.pone.0090877https://doaj.org/article/706fdac6606d4fa8a841e3921766d00f2014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24595403/?tool=EBIhttps://doaj.org/toc/1932-6203We 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.Faming TianMarta GreplováIvo FrébortNicholas DaleRichard NapierPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 3, p e90877 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Faming Tian Marta Greplová Ivo Frébort Nicholas Dale Richard Napier A highly selective biosensor with nanomolar sensitivity based on cytokinin dehydrogenase.
description	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.
format	article
author	Faming Tian Marta Greplová Ivo Frébort Nicholas Dale Richard Napier
author_facet	Faming Tian Marta Greplová Ivo Frébort Nicholas Dale Richard Napier
author_sort	Faming Tian
title	A highly selective biosensor with nanomolar sensitivity based on cytokinin dehydrogenase.
title_short	A highly selective biosensor with nanomolar sensitivity based on cytokinin dehydrogenase.
title_full	A highly selective biosensor with nanomolar sensitivity based on cytokinin dehydrogenase.
title_fullStr	A highly selective biosensor with nanomolar sensitivity based on cytokinin dehydrogenase.
title_full_unstemmed	A highly selective biosensor with nanomolar sensitivity based on cytokinin dehydrogenase.
title_sort	highly selective biosensor with nanomolar sensitivity based on cytokinin dehydrogenase.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/706fdac6606d4fa8a841e3921766d00f
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A highly selective biosensor with nanomolar sensitivity based on cytokinin dehydrogenase.

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