Determination of haloalkane dehalogenase activity by capillary zone electrophoresis

Zdeněk Glatz, Mária V. Marini, Michaela Wimmerová, Jiří Damborský, Yuji Nagata

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)

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 monitored by sequential capillary zone electrophoresis runs. The determinations were performed in a 75 μm 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 β-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 both cases 25°C. The method is rapid, can be automated, and requires only small amount of enzyme preparation and substrate. (C) 2000 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)219-225
Number of pages7
JournalJournal of Chromatography A
Volume895
Issue number1-2
DOIs
Publication statusPublished - 2000 Oct 20

Keywords

  • Enzymes
  • Haloalkane dehalogenase
  • Haloalkanes

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

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