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 a Michaela WIMMEROVÁ

Základní údaje

Originální název

Determination of haloalkane dehalogenase enzymatic activity by capillary zone electrophoresis

Autoři

GLATZ, Zdeněk (203 Česká republika, garant), María Victoria MARINI PALOMEQUE (858 Uruguay) a Michaela WIMMEROVÁ (203 Česká republika)

Vydání

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

Nakladatel

Universites des Saarlandes, Germany

Další údaje

Jazyk

angličtina

Typ výsledku

Stať ve sborníku

Obor

10600 1.6 Biological sciences

Stát vydavatele

Německo

Utajení

není předmětem státního či obchodního tajemství

Kód RIV

RIV/00216224:14310/00:00003266

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

haloalkane dehalogenase; enzymatic activity; capillary zone electrophoresis

Štítky

Capillary Zone Electrophoresis, enzymatic activity, haloalkane dehalogenase

Změněno: 14. 11. 2008 18:01, Mgr. María Victoria Marini Palomeque, Ph.D.

Anotace

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.

Návaznosti

GA203/97/P149, projekt VaV

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

Investor: Grantová agentura ČR, Studium molekulárních mechanismů biodegradačních reakcí - konstrukce QSBR modelů a proteinové inženýrství dehalogenas

Citovat

GLATZ, Zdeněk, María Victoria MARINI PALOMEQUE a 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, s. 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 a 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, s.~191.

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