An Ultrasensitive Fluorescence Assay for the Detection of
Halides and Enzymatic Dehalogenation

J 2020

An Ultrasensitive Fluorescence Assay for the Detection of Halides and Enzymatic Dehalogenation

ASLAN-UZEL, Askin S., Andy BEIER, David KOVÁŘ, Clemens CZIEGLER, Santosh K. PADHI et. al.

Základní údaje

Originální název

An Ultrasensitive Fluorescence Assay for the Detection of Halides and Enzymatic Dehalogenation

Autoři

ASLAN-UZEL, Askin S. (276 Německo), Andy BEIER (276 Německo, domácí), David KOVÁŘ (203 Česká republika, domácí), Clemens CZIEGLER (40 Rakousko), Santosh K. PADHI (356 Indie), Eva D. SCHUITEN (276 Německo), Mark DORR (276 Německo), Dominique BOTTCHER (276 Německo), Frank HOLLMANN (528 Nizozemské království), Florian RUDROFF (40 Rakousko), Marko D. MIHOVILOVIC (40 Rakousko), Tomáš BURYŠKA (203 Česká republika, domácí), Jiří DAMBORSKÝ (203 Česká republika, garant, domácí), Zbyněk PROKOP (203 Česká republika, domácí), Christoffel P. S. BADENHORST (276 Německo) a Uwe T. BORNSCHEUER (276 Německo)

Vydání

ChemCatChem, WEINHEIM, WILEY-V C H VERLAG GMBH, 2020, 1867-3880

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10403 Physical chemistry

Stát vydavatele

Německo

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 5.686

Kód RIV

RIV/00216224:14310/20:00115467

Organizační jednotka

Přírodovědecká fakulta

DOI

http://dx.doi.org/10.1002/cctc.201901891

UT WoS

000510215900001

Klíčová slova anglicky

dehalogenase; fluorescence; halides; haloalkane; haloperoxidase

Štítky

rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 16. 2. 2023 12:32, Mgr. Michaela Hylsová, Ph.D.

Anotace

V originále

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.

Návaznosti

EF16_013/0001761, projekt VaV

Název: RECETOX RI

EF19_074/0012727, projekt VaV

Název: MSCAfellow3@MUNI

LM2015051, projekt VaV

Název: Centrum pro výzkum toxických látek v prostředí (Akronym: RECETOX RI)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Výzkumná infrastruktura RECETOX

Citovat

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 a Uwe T. BORNSCHEUER. An Ultrasensitive Fluorescence Assay for the Detection of Halides and Enzymatic Dehalogenation. ChemCatChem. WEINHEIM: WILEY-V C H VERLAG GMBH, 2020, roč. 12, č. 7, s. 2032-2039. ISSN 1867-3880. Dostupné z: https://dx.doi.org/10.1002/cctc.201901891.

@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 a 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, roč.~12, č.~7, s.~2032-2039. ISSN~1867-3880. Dostupné z: https://dx.doi.org/10.1002/cctc.201901891.

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