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. p. 191. 2000.
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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
Original language English
Type of outcome Proceedings paper
Field of Study 10600 1.6 Biological sciences
Country of publisher Germany
Confidentiality degree is not subject to a state or trade secret
RIV identification code RIV/00216224:14310/00:00003266
Organization unit Faculty of Science
Keywords in English haloalkane dehalogenase; enzymatic activity; capillary zone electrophoresis
Tags Capillary Zone Electrophoresis, enzymatic activity, haloalkane dehalogenase
Changed by Changed by: Mgr. María Victoria Marini Palomeque, Ph.D., učo 22912. Changed: 14/11/2008 18:01.
Abstract
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 projectName: 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
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