Application of capillary zone electrophoresis for the study and
assay of enzymes

D 2002

Application of capillary zone electrophoresis for the study and assay of enzymes

NOVÁKOVÁ, Soňa and Zdeněk GLATZ

Basic information

Original name

Application of capillary zone electrophoresis for the study and assay of enzymes

Authors

NOVÁKOVÁ, Soňa and Zdeněk GLATZ

Edition

2002. vyd. Brno, Book of abstracts of 6th Meeting of biochemists and molecular biologists, p. 77-77, 2002

Publisher

MU Brno

Other information

Language

English

Type of outcome

Stať ve sborníku

Field of Study

10600 1.6 Biological sciences

Country of publisher

Czech Republic

Confidentiality degree

není předmětem státního či obchodního tajemství

Organization unit

Faculty of Science

Keywords in English

EMMA; enzymes

Abstract

V originále

Ten years ago a new application for the evaluation of enzymatic reactions in capillary electrophoresis was proposed and developed, electrophoretically mediated microanalysis (EMMA). In this method, 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. Since its discovery by Bao and Regnier the EMMA methodology has been utilized in a number of biochemical systems: for assays of enzyme activities, determinations substrates, Michaelis constants, inhibitors and inhibition constants, etc. In this communication the combination of the EMMA methodology with a partial filling technique was applied to study the kinetic parameters of bi-substrate enzymatic reaction of rhodanese. In this set-up the part of the capillary is filled with the buffer best for the enzymatic reaction whereas the rest of the capillary with the background electrolyte optimal for separation of substrates and products. The basic limitation of EMMA methodology the necessity to have the electrophoretic conditions compatible with both the separation of substrate(s) and product(s) of the enzymatic reaction and the enzymatic reaction is thus overcome.

Links

GA525/00/0785, research and development project

Name: Acidifikační procesy v sulfidových odpadech

Investor: Czech Science Foundation, Acidification processes in sulfide wastes

Citovat

NOVÁKOVÁ, Soňa and Zdeněk GLATZ. Application of capillary zone electrophoresis for the study and assay of enzymes. In Book of abstracts of 6th Meeting of biochemists and molecular biologists. 2002nd ed. Brno: MU Brno, 2002, p. 77.

@inproceedings{403280,
   author = {Nováková, Soňa and Glatz, Zdeněk},
   address = {Brno},
   booktitle = {Book of abstracts of 6th Meeting of biochemists and molecular biologists},
   edition = {2002},
   keywords = {EMMA; enzymes},
   language = {eng},
   location = {Brno},
   pages = {77-77},
   publisher = {MU Brno},
   title = {Application of capillary zone electrophoresis for the study and assay of enzymes},
   year = {2002}
}

TY  - JOUR
ID  - 403280
AU  - Nováková, Soňa - Glatz, Zdeněk
PY  - 2002
TI  - Application of capillary zone electrophoresis for the study and assay of enzymes
PB  - MU Brno
CY  - Brno
KW  - EMMA
KW  - enzymes
N2  - Ten years ago a new application for the evaluation of enzymatic reactions in capillary electrophoresis was proposed and developed, electrophoretically mediated microanalysis (EMMA). In this method, 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. Since its discovery by Bao and Regnier the EMMA methodology has been utilized in a number of biochemical systems: for assays of enzyme activities, determinations substrates, Michaelis constants, inhibitors and inhibition constants, etc. In this communication the combination of the EMMA methodology with a partial filling technique was applied to study the kinetic parameters of bi-substrate enzymatic reaction of rhodanese. In this set-up the part of the capillary is filled with the buffer best for the enzymatic reaction whereas the rest of the capillary with the background electrolyte optimal for separation of substrates and products. The basic limitation of EMMA methodology the necessity to have the electrophoretic conditions compatible with both the separation of substrate(s) and product(s) of the enzymatic reaction and the enzymatic reaction is thus overcome.
ER  -

NOVÁKOVÁ, Soňa and Zdeněk GLATZ. Application of capillary zone electrophoresis for the study and assay of enzymes. In \textit{Book of abstracts of 6th Meeting of biochemists and molecular biologists}. 2002nd ed. Brno: MU Brno, 2002, p.~77.

Detailed Information on Publication Record