D 2002

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

NOVÁKOVÁ, Soňa a Zdeněk GLATZ

Základní údaje

Originální název

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

Vydání

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

Nakladatel

MU Brno

Další údaje

Jazyk

angličtina

Typ výsledku

Stať ve sborníku

Obor

10600 1.6 Biological sciences

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

EMMA; enzymes

Štítky

Změněno: 19. 5. 2009 18:44, prof. RNDr. Zdeněk Glatz, CSc.

Anotace

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.

Návaznosti

GA525/00/0785, projekt VaV
Název: Acidifikační procesy v sulfidových odpadech
Investor: Grantová agentura ČR, Acidifikační procesy v sulfidových odpadech