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Capillary electrophoresis as a tool for a cost‐effective assessment of the activity of plant membrane enzyme chlorophyllase
Author(s) -
Nowak Paweł,
Michalik Maciej,
Fiedor Leszek,
Woźniakiewicz Michał,
Kościelniak Paweł
Publication year - 2013
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201300304
Subject(s) - chromatography , reagent , capillary electrophoresis , chemistry , membrane , capillary action , enzyme , enzyme assay , electrophoresis , biochemistry , materials science , composite material
The potential of the CE‐based enzymatic assay has been demonstrated in case of a typical plant membrane enzyme – chlorophyllase. An efficient, automated and rapid semi‐quantitative method has been developed, which allowed us to assess the activity of the enzyme via two strategies. Firstly, a reaction conducted in a vial placed directly on the sample tray was combined with the concomitant separation and detection of reagents. The method was used to monitor the reaction progress. Secondly, an online approach was applied using an electrophoretically mediated mixing. The reaction was performed in‑capillary, resulting in an extreme reduction of the reagent amounts required for a single run. Both methods were effective in the assessment of the activity of a membrane enzyme, a member of protein class known to pose experimental difficulties.