title: Capillary electrophoresis as a tool for a cost-effective assessment of the activity of plant membrane enzyme chlorophyllase.
abstract: The potential of the capillary electrophoresis-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 on-line approach was applied by 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.
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemistry, Jagiellonian University, Krak ów, Poland.
authors.forename: Paweł
authors.initials: P
authors.lastname: Nowak
authors.forename: Maciej
authors.initials: M
authors.lastname: Michalik
authors.forename: Leszek
authors.initials: L
authors.lastname: Fiedor
authors.forename: Michał
authors.initials: M
authors.lastname: Woźniakiewicz
authors.forename: Paweł
authors.initials: P
authors.lastname: Kościelniak
mesh_terms: D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D002734:Chlorophyll / Q000378:metabolism; D000077194:Chlorophyll A; D019075:Electrophoresis, Capillary / Q000191:economics / Q000379:methods*; D057075:Enzyme Assays / Q000191:economics / Q000379:methods*; D018515:Plant Leaves / Q000201:enzymology / Q000378:metabolism; D018724:Spinacia oleracea / Q000201:enzymology* / Q000378:metabolism
chemical_list: D002734:Chlorophyll; D002265:Carboxylic Ester Hydrolases; C019875:chlorophyllase; D000077194:Chlorophyll A