Determination of the Kinetic Parameters of Rhodanese by Electrophoretically Mediated Microanalysis in a Partially Filled Capillary
| Autoři | |
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| Rok publikování | 2001 |
| Druh | Článek ve sborníku |
| Konference | Book of abstract 2nd International symposium "Separations in BioSciences SBS 2001" |
| Fakulta / Pracoviště MU | |
| Citace | |
| Obor | Biochemie |
| Klíčová slova | EMMA rhodanese |
| Popis | Recently, a new application for the evaluation of enzymatic reactions in capillary electrophoresis 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 communication the EMMA technique in was applied to determine the kinetic parameters of rhodanese - temperature optimum, KM for thiosulfate and cyanide and type of kinetic mechanism of enzymatic reaction. 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: S2O32- + CN- Ąú SCN- + SO32- 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. |
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