Determination of haloalkane dehalogenase enzymatic activity by
capillary zone electrophoresis

D 2000

Determination of haloalkane dehalogenase enzymatic activity by capillary zone electrophoresis

GLATZ, Zdeněk, María Victoria MARINI PALOMEQUE and Michaela WIMMEROVÁ

Basic information

Original name

Determination of haloalkane dehalogenase enzymatic activity by capillary zone electrophoresis

Authors

GLATZ, Zdeněk (203 Czech Republic, guarantor), María Victoria MARINI PALOMEQUE (858 Uruguay) and Michaela WIMMEROVÁ (203 Czech Republic)

Edition

Saarbrucken, Germany, Book of Abstract "13th International symposium on high performance capillary electrophoresis and related techniques HPCE 2000". p. 191-191, 2000

Publisher

Universites des Saarlandes, Germany

Other information

Language

English

Type of outcome

Stať ve sborníku

Field of Study

10600 1.6 Biological sciences

Country of publisher

Germany

Confidentiality degree

není předmětem státního či obchodního tajemství

RIV identification code

RIV/00216224:14310/00:00003266

Organization unit

Faculty of Science

Keywords in English

haloalkane dehalogenase; enzymatic activity; capillary zone electrophoresis

Abstract

V originále

A new sensitive method has been developed for the determination of haloalkane dehalogenase activity. The enzymatic reactions were carried out directly in thermostatted autosampler vials and the formation of product - bromide or chloride ions - was monito red by sequential capillary zone electrophoresis runs. The determinations were performed in a 75 mm fused-silica capillary using 5 mM chromate, 0.5 mM tetradecyltrimethylammonium bromide (pH 8.4) as a background electrolyte, separation voltage 15 kV (negative polarity ) and indirect detection at sample wavelength 315 nm, reference wavelength 375 nm for brominated and chlorinated substrates, respectively 0.1 M b-alanine-HCl (pH 3.50) as a background electrolyte, separation voltage 18 kV (negative polarity) and direct detection at 200 nm for brominated substrates. The temperature of capillary was in bot cases 25oC. The method is rapid, can be automated, and requires only small amount of enzyme preparation and substrate.

Links

GA203/97/P149, research and development project

Name: Studium molekulárních mechanismů biodegradačních reakcí - konstrukce QSBR modelů a proteinové inženýrství dehalogenas

Investor: Czech Science Foundation, Study of the molecular mechanisms of biodegradation reactions - construction of QSBR models and protein engineering of haloalkane dehalogenases

Citovat

GLATZ, Zdeněk, María Victoria MARINI PALOMEQUE and Michaela WIMMEROVÁ. Determination of haloalkane dehalogenase enzymatic activity by capillary zone electrophoresis. In Book of Abstract "13th International symposium on high performance capillary electrophoresis and related techniques HPCE 2000". Saarbrucken, Germany: Universites des Saarlandes, Germany, 2000, p. 191.

@inproceedings{243631,
   author = {Glatz, Zdeněk and Marini Palomeque, María Victoria and Wimmerová, Michaela},
   address = {Saarbrucken, Germany},
   booktitle = {Book of Abstract "13th International symposium on high performance capillary electrophoresis and related techniques HPCE 2000".},
   keywords = {haloalkane dehalogenase; enzymatic activity; capillary zone electrophoresis},
   language = {eng},
   location = {Saarbrucken, Germany},
   pages = {191-191},
   publisher = {Universites des Saarlandes, Germany},
   title = {Determination of haloalkane dehalogenase enzymatic activity by capillary zone electrophoresis},
   year = {2000}
}

TY  - JOUR
ID  - 243631
AU  - Glatz, Zdeněk - Marini Palomeque, María Victoria - Wimmerová, Michaela
PY  - 2000
TI  - Determination of haloalkane dehalogenase enzymatic activity by capillary zone electrophoresis
PB  - Universites des Saarlandes, Germany
CY  - Saarbrucken, Germany
KW  - haloalkane dehalogenase
KW  - enzymatic activity
KW  - capillary zone electrophoresis
N2  - A new sensitive method has been developed for the determination of haloalkane dehalogenase activity. The enzymatic reactions were carried out directly in thermostatted autosampler vials and the formation of product - bromide or chloride ions - was monito red by sequential capillary zone electrophoresis runs. The determinations were performed in a 75 mm fused-silica capillary using 5 mM chromate, 0.5 mM tetradecyltrimethylammonium bromide (pH 8.4) as a background electrolyte, separation voltage 15 kV (negative polarity ) and indirect detection at sample wavelength 315 nm, reference wavelength 375 nm for brominated and chlorinated substrates, respectively 0.1 M b-alanine-HCl (pH 3.50) as a background electrolyte, separation voltage 18 kV (negative polarity) and direct detection at 200 nm for brominated substrates. The temperature of capillary was in bot cases 25oC. The method is rapid, can be automated, and requires only small amount of enzyme preparation and substrate.
ER  -

GLATZ, Zdeněk, María Victoria MARINI PALOMEQUE and Michaela WIMMEROVÁ. Determination of haloalkane dehalogenase enzymatic activity by capillary zone electrophoresis. In \textit{Book of Abstract ''13th International symposium on high performance capillary electrophoresis and related techniques HPCE 2000''.}. Saarbrucken, Germany: Universites des Saarlandes, Germany, 2000, p.~191.

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