| ASLAN-UZEL, Askin S., Andy BEIER, David KOVÁŘ, Clemens CZIEGLER, Santosh K. PADHI, Eva D. SCHUITEN, Mark DORR, Dominique BOTTCHER, Frank HOLLMANN, Florian RUDROFF, Marko D. MIHOVILOVIC, Tomáš BURYŠKA, Jiří DAMBORSKÝ, Zbyněk PROKOP, Christoffel P. S. BADENHORST and Uwe T. BORNSCHEUER. An Ultrasensitive Fluorescence Assay for the Detection of Halides and Enzymatic Dehalogenation. ChemCatChem. WEINHEIM: WILEY-V C H VERLAG GMBH, 2020, vol. 12, No 7, p. 2032-2039. ISSN 1867-3880. Available from: https://dx.doi.org/10.1002/cctc.201901891. |
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@article{1640717,
author = {AslanandUzel, Askin S. and Beier, Andy and Kovář, David and Cziegler, Clemens and Padhi, Santosh K. and Schuiten, Eva D. and Dorr, Mark and Bottcher, Dominique and Hollmann, Frank and Rudroff, Florian and Mihovilovic, Marko D. and Buryška, Tomáš and Damborský, Jiří and Prokop, Zbyněk and Badenhorst, Christoffel P. S. and Bornscheuer, Uwe T.},
article_location = {WEINHEIM},
article_number = {7},
doi = {http://dx.doi.org/10.1002/cctc.201901891},
keywords = {dehalogenase; fluorescence; halides; haloalkane; haloperoxidase},
language = {eng},
issn = {1867-3880},
journal = {ChemCatChem},
title = {An Ultrasensitive Fluorescence Assay for the Detection of Halides and Enzymatic Dehalogenation},
url = {https://onlinelibrary.wiley.com/doi/full/10.1002/cctc.201901891},
volume = {12},
year = {2020}
}
TY - JOUR
ID - 1640717
AU - Aslan-Uzel, Askin S. - Beier, Andy - Kovář, David - Cziegler, Clemens - Padhi, Santosh K. - Schuiten, Eva D. - Dorr, Mark - Bottcher, Dominique - Hollmann, Frank - Rudroff, Florian - Mihovilovic, Marko D. - Buryška, Tomáš - Damborský, Jiří - Prokop, Zbyněk - Badenhorst, Christoffel P. S. - Bornscheuer, Uwe T.
PY - 2020
TI - An Ultrasensitive Fluorescence Assay for the Detection of Halides and Enzymatic Dehalogenation
JF - ChemCatChem
VL - 12
IS - 7
SP - 2032-2039
EP - 2032-2039
PB - WILEY-V C H VERLAG GMBH
SN - 18673880
KW - dehalogenase
KW - fluorescence
KW - halides
KW - haloalkane
KW - haloperoxidase
UR - https://onlinelibrary.wiley.com/doi/full/10.1002/cctc.201901891
L2 - https://onlinelibrary.wiley.com/doi/full/10.1002/cctc.201901891
N2 - Halide assays are important for the study of enzymatic dehalogenation, a topic of great industrial and scientific importance. Here we describe the development of a very sensitive halide assay that can detect less than a picomole of bromide ions, making it very useful for quantifying enzymatic dehalogenation products. Halides are oxidised under mild conditions using the vanadium-dependent chloroperoxidase from Curvularia inaequalis, forming hypohalous acids that are detected using aminophenyl fluorescein. The assay is up to three orders of magnitude more sensitive than currently available alternatives, with detection limits of 20 nM for bromide and 1 mu M for chloride and iodide. We demonstrate that the assay can be used to determine specific activities of dehalogenases and validate this by comparison to a well-established GC-MS method. This new assay will facilitate the identification and characterisation of novel dehalogenases and may also be of interest to those studying other halide-producing enzymes.
ER -
ASLAN-UZEL, Askin S., Andy BEIER, David KOVÁŘ, Clemens CZIEGLER, Santosh K. PADHI, Eva D. SCHUITEN, Mark DORR, Dominique BOTTCHER, Frank HOLLMANN, Florian RUDROFF, Marko D. MIHOVILOVIC, Tomáš BURYŠKA, Jiří DAMBORSKÝ, Zbyněk PROKOP, Christoffel P. S. BADENHORST and Uwe T. BORNSCHEUER. An Ultrasensitive Fluorescence Assay for the Detection of Halides and Enzymatic Dehalogenation. \textit{ChemCatChem}. WEINHEIM: WILEY-V C H VERLAG GMBH, 2020, vol.~12, No~7, p.~2032-2039. ISSN~1867-3880. Available from: https://dx.doi.org/10.1002/cctc.201901891.
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