Stanovení kinetických parametrů rhodanasy pomoci metody EMMA (Electrophoretivally Mediated Microanalysis)
| Autoři | |
|---|---|
| Rok publikování | 2001 |
| Druh | Článek ve sborníku |
| Konference | Sborník příspěvků 5.Pracovního setkání biochemiků a molekulárních biologů |
| Fakulta / Pracoviště MU | |
| Citace | |
| Obor | Biochemie |
| Klíčová slova | Rhodanese; Electrophoretivally Mediated Microanalysis |
| Popis | Recently, a new application for the evaluation of enzymatic reactions in capillary electrophoresi was proposed, electrophoretically mediated microanalysis (EMMA). In this technique, 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, the 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 paper the EMMA technique was applied to determine the kinetic parameters of rhodanese. Rhodanese (thiosulfate: cyanide sulfur transferase, EC 2.8.1.1) was discovered in 1933 by Lang. It is responsible for the transfer of the sulfane sulfur of thiosulfate to an acceptor, which is likely to be cyanide under some physiological conditions. Rhodaneses are practically ubiquitous enzymes, their activity has been detected in several species ranging from microorganisms through fungi, plants and animals to man. In bacteria the enzyme was found in a variety of heterotrophic, photolithotrophic species as well as certain chemolithotrophic organisms including Thiobacillus sp. To the best of our knowledge, the EMMA has not yet been applied for two substrate enzymatic reaction. |
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