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.
Highlights
We demonstrate that the assay can be used to determine specific activities of dehalogenases and validate this by comparison to a wellestablished gas chromatography and mass spectrometry (GC-MS) method
We demonstrate that the new halide assay is up to three orders of magnitude more sensitive than currently available alternatives and validated it by comparison to a standard GC-MS method for determining the specific activities of two model dehalogenases
The two distinct topics discussed in this paper, haloalkane dehalogenases and halide assays, are united by the unmet demand for more powerful dehalogenase assays.[10]
Summary
S., Beier, A., Kovář, D., Cziegler, C., Padhi, S. D., Dörr, M., Böttcher, D., Hollmann, F., & More Authors (2019). An Ultrasensitive Fluorescence Assay for the Detection of Halides and Enzymatic Dehalogenation.
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