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

Kód výsledku v 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>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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

Název v anglickém jazyce

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

Popis výsledku anglicky

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

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

CE - Biochemie

OECD FORD obor

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Projekt

<a href="/cs/project/GA525%2F00%2F0785" target="_blank" >GA525/00/0785: Acidifikační procesy v sulfidových odpadech</a><br>

Návaznosti

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

Rok uplatnění

2002

Kód důvěrnosti údajů

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

Název periodika

Electrophoresis

ISSN

0173-0835

e-ISSN

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Svazek periodika

23

Číslo periodika v rámci svazku

7-8

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

7

Strana od-do

1063

Kód UT WoS článku

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EID výsledku v databázi Scopus

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