2006
Electrophoretically mediated microanalysis as a tool for the enzyme study
TELNAROVÁ, Magdaléna, Kateřina PAPEŽOVÁ, Tomáš NĚMEC a Zdeněk GLATZZákladní údaje
Originální název
Electrophoretically mediated microanalysis as a tool for the enzyme study
Název česky
Studium enzymů pomocí metody EMMA
Autoři
TELNAROVÁ, Magdaléna, Kateřina PAPEŽOVÁ, Tomáš NĚMEC a Zdeněk GLATZ
Vydání
Belgium, CD of the abstracts of the 29th international symposium on capillary chromatography, od s. x, 1 s. 2006
Nakladatel
T. Sandra and P. Sandra
Další údaje
Jazyk
angličtina
Typ výsledku
Stať ve sborníku
Obor
10600 1.6 Biological sciences
Stát vydavatele
Itálie
Utajení
není předmětem státního či obchodního tajemství
Organizační jednotka
Přírodovědecká fakulta
Klíčová slova anglicky
cytochromes P450; diclofenac; MEKC
Štítky
Změněno: 19. 5. 2009 18:44, prof. RNDr. Zdeněk Glatz, CSc.
V originále
Fourteen years ago a new application for the evaluation of enzymatic reactions in capillary electrophoresis (CE) was proposed and developed, electrophoretically mediated microanalysis (EMMA). In this methodology, the capillary is used not only as a separation medium but also as a reaction chamber. Substrate(s) and enzyme are introduced in the capillary as distinct plugs, the first analyte injected being the one with the lower electrophoretic mobility. Upon the application of an electric field, these two zones interpenetrate due the differences in their electrophoretic mobilities. Enzymatic reaction takes place and the resultant reaction product(s) and the unreacted substrate(s) are electrophoretically transported towards detector, where they are individually detected. In this communication the EMMA methodology was applied to complex kinetic study of haloalkane dehalogenase from Sphingomonas paucimobilis UT26. The Michaelis constants for different substrates, the inhibition constant for inhibitor 1,2-dichloroethane, substrate inhibition of 1,2-dibromoethane and the effect of temperature on enzymatic reaction were evaluated by means of the EMMA methodology with partial filling technique. EMMA combines all advantages of CE in the study of enzyme kinetics. The minimal sample and reagent requirements as well as high separation efficiency of CE are fully utilŹized. The absence of manual procedures and complete automation miniŹmize the risk of cross contamination and strongly reduce the assay cost.
Česky
V píspvku byla prezentována možnost využití metody EMMA pi studiu enzym.
Návaznosti
| GA203/06/0047, projekt VaV |
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| LC06023, projekt VaV |
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| MSM0021622413, záměr |
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