TY  - JOUR
U1  - Wissenschaftlicher Artikel
A1  - Aslan‐Üzel, Aşkın S.
A1  - Beier, Andy
A1  - Kovář, David
A1  - Cziegler, Clemens
A1  - Padhi, Santosh K.
A1  - Schuiten, Eva D.
A1  - Dörr, Mark
A1  - Böttcher, Dominique
A1  - Hollmann, Frank
A1  - Rudroff, Florian
A1  - Mihovilovic, Marko D.
A1  - Buryška, Tomáš
A1  - Damborský, Jiří
A1  - Prokop, Zbyněk
A1  - Badenhorst, Christoffel P. S.
A1  - Bornscheuer, Uwe T.
T1  - An Ultrasensitive Fluorescence Assay for the Detection of Halides and Enzymatic Dehalogenation
JF  - ChemCatChem
N2  - Abstract
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 μM 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.
KW  - -
KW  - dehalogenase
KW  - fluorescence
KW  - halides
KW  - haloalkane
KW  - haloperoxidase
Y1  - 2020
UN  - https://nbn-resolving.org/urn:nbn:de:gbv:9-opus-40008
U6  - https://doi.org/10.1002/cctc.201901891
DO  - https://doi.org/10.1002/cctc.201901891
VL  - 12
IS  - 7
SP  - 2032
EP  - 2039
PB  - Wiley
CY  - Hoboken, New Jersey
ER  -