Determination of the kinetic parameters of rhodanese by electrophoretically mediated microanalysis in a partially filled capillary

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F02%3A00005658" target="_blank" >RIV/00216224:14310/02:00005658 - isvavai.cz</a>

Result on the web

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DOI - Digital Object Identifier

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Result language

angličtina

Original language name

Determination of the kinetic parameters of rhodanese by electrophoretically mediated microanalysis in a partially filled capillary

Original language description

This report describes the application of electrophoretically mediated microanalysis (EMMA) for the study of kinetic parameters of bi-substrate enzymatic reaction of rhodanese. The Michaelis constants (Km) for both substrates and the effect of temperatureon rhodanese reaction were evaluated by means of the combination of the EMMA methodology with a partial filling technique. 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 enzymatic reaction was performed in 25 mM HEPES buffer (pH 8.5) while the low pH background electrolyte 100 mM beta-alanine - HCl (pH 3.5) was used for separation of substrates and products that are the inorganic anions. The estimated value of Km for thiosulfate of 1.30 x 10-2 M was consistent with previously published values; the Km for cyanide of 7.6 x 10-3 M was determined for the first time. In

Czech name

Determination of the kinetic parameters of rhodanese by electrophoretically mediated microanalysis in a partially filled capillary

Czech description

This report describes the application of electrophoretically mediated microanalysis (EMMA) for the study of kinetic parameters of bi-substrate enzymatic reaction of rhodanese. The Michaelis constants (Km) for both substrates and the effect of temperatureon rhodanese reaction were evaluated by means of the combination of the EMMA methodology with a partial filling technique. 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 enzymatic reaction was performed in 25 mM HEPES buffer (pH 8.5) while the low pH background electrolyte 100 mM beta-alanine - HCl (pH 3.5) was used for separation of substrates and products that are the inorganic anions. The estimated value of Km for thiosulfate of 1.30 x 10-2 M was consistent with previously published values; the Km for cyanide of 7.6 x 10-3 M was determined for the first time. In

Type

J_x - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)

CEP classification

CE - Biochemistry

OECD FORD branch

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Project

<a href="/en/project/GA525%2F00%2F0785" target="_blank" >GA525/00/0785: Acidification processes in sulphide wastes</a><br>

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2002

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Electrophoresis

ISSN

0173-0835

e-ISSN

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Volume of the periodical

23

Issue of the periodical within the volume

7-8

Country of publishing house

US - UNITED STATES

Number of pages

7

Pages from-to

1063

UT code for WoS article

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EID of the result in the Scopus database

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