Electrophoretically mediated microanalysis as a tool for the
enzyme study

D 2006

Electrophoretically mediated microanalysis as a tool for the enzyme study

TELNAROVÁ, Magdaléna, Kateřina PAPEŽOVÁ, Tomáš NĚMEC and Zdeněk GLATZ

Basic information

Original name

Electrophoretically mediated microanalysis as a tool for the enzyme study

Name in Czech

Studium enzymů pomocí metody EMMA

Authors

TELNAROVÁ, Magdaléna, Kateřina PAPEŽOVÁ, Tomáš NĚMEC and Zdeněk GLATZ

Edition

Belgium, CD of the abstracts of the 29th international symposium on capillary chromatography, p. x, 1 pp. 2006

Publisher

T. Sandra and P. Sandra

Other information

Language

English

Type of outcome

Stať ve sborníku

Field of Study

10600 1.6 Biological sciences

Country of publisher

Italy

Confidentiality degree

není předmětem státního či obchodního tajemství

Organization unit

Faculty of Science

Keywords in English

cytochromes P450; diclofenac; MEKC

Abstract

ORIG CZ

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.

In Czech

V píspvku byla prezentována možnost využití metody EMMA pi studiu enzym.

Links

GA203/06/0047, research and development project

Name: Využití kapilární elektroforézy pro studium metabolismu léčiv

Investor: Czech Science Foundation, Capillary electrophoresis as a tool for the drug-metabolism studies

LC06023, research and development project

Name: Integrované bioanalytické technologie pro mikroanalýzy a diagnostiku s využitím LIF a hmotnostní spektrometrie

Investor: Ministry of Education, Youth and Sports of the CR

MSM0021622413, plan (intention)

Name: Proteiny v metabolismu a při interakci organismů s prostředím

Investor: Ministry of Education, Youth and Sports of the CR, Proteins in metabolism and interaction of organisms with the environment

Citovat

TELNAROVÁ, Magdaléna, Kateřina PAPEŽOVÁ, Tomáš NĚMEC and Zdeněk GLATZ. Electrophoretically mediated microanalysis as a tool for the enzyme study. In CD of the abstracts of the 29th international symposium on capillary chromatography. Belgium: T. Sandra and P. Sandra, 2006, p. x, 1 pp.

@inproceedings{636991,
   author = {Telnarová, Magdaléna and Papežová, Kateřina and Němec, Tomáš and Glatz, Zdeněk},
   address = {Belgium},
   booktitle = {CD of the abstracts of the 29th international symposium on capillary chromatography},
   keywords = {cytochromes P450; diclofenac; MEKC},
   language = {eng},
   location = {Belgium},
   pages = {x},
   publisher = {T. Sandra and P. Sandra},
   title = {Electrophoretically mediated microanalysis as a tool for the enzyme study},
   year = {2006}
}

TY  - JOUR
ID  - 636991
AU  - Telnarová, Magdaléna - Papežová, Kateřina - Němec, Tomáš - Glatz, Zdeněk
PY  - 2006
TI  - Electrophoretically mediated microanalysis as a tool for the enzyme study
PB  - T. Sandra and P. Sandra
CY  - Belgium
KW  - cytochromes P450
KW  - diclofenac
KW  - MEKC
N2  - 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.
ER  -

TELNAROVÁ, Magdaléna, Kateřina PAPEŽOVÁ, Tomáš NĚMEC and Zdeněk GLATZ. Electrophoretically mediated microanalysis as a tool for the enzyme study. In \textit{CD of the abstracts of the 29th international symposium on capillary chromatography}. Belgium: T. Sandra and P. Sandra, 2006, p.~x, 1 pp.

Detailed Information on Publication Record