Monitoring tripeptidase activity using capillary electrophoresis
| Autor: | Gordon R. Hague, Francis Mulholland, Takafumi Kasumi, Sara Movahedi |
|---|---|
| Rok vydání: | 1993 |
| Předmět: |
Gel electrophoresis
Chromatography biology Chemistry Organic Chemistry Lactococcus lactis General Medicine biology.organism_classification Biochemistry Tripeptidase activity Analytical Chemistry Standard curve chemistry.chemical_compound Capillary electrophoresis Ninhydrin Enzyme kinetics Derivatization |
| Zdroj: | Journal of Chromatography A. 636:63-68 |
| ISSN: | 0021-9673 |
| DOI: | 10.1016/0021-9673(93)80057-f |
| Popis: | Capillary electrophoresis (CE) was used to assay the activity of a tripeptidase from a crude extract of Lactococcus lactis subsp. lactis NCDO 712 against the substrate, Gly—Gly—Phe and a comparison with a standard ninhydrin assay was made. Standard curves of the substrates and products showed a significantly variable colorimetric reaction to ninhydrin making accurate quantification of the tripeptidase problematic. The CE assay further demonstrated that the presence of contaminating enzymes in crude cell-free extracts can cause secondary reactions that are not apparent from the ninhydrin assay data. The CE assay was also able to generate enzyme kinetics data and monitor, during purification, the presence of co-eluting contaminating activities. The speed and sensitivity with CE allows routine analysis of the tripeptidase activity without any derivatization normally required for this enzyme. |
| Databáze: | OpenAIRE |
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