Purification of α-glucosidase from mouse intestine by countercurrent chromatography coupled with a reverse micelle solvent system
| Autor: | Xiaoli Ye, Yinran Hu, Yubo Xiao, Yong Yang, Zongyao Zou, Kai He, Xuegang Li, Pincao Gao |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Chromatography Sodium 010401 analytical chemistry chemistry.chemical_element Filtration and Separation 01 natural sciences Micelle 0104 chemical sciences Analytical Chemistry Solvent 03 medical and health sciences 030104 developmental biology Countercurrent chromatography chemistry Pulmonary surfactant Phase (matter) Protein purification Polyacrylamide gel electrophoresis |
| Zdroj: | Journal of Separation Science. 39:703-708 |
| ISSN: | 1615-9306 |
| DOI: | 10.1002/jssc.201501092 |
| Popis: | Countercurrent chromatography coupled with a reverse micelle solvent was applied to separate α-glucosidase, which is stable at pH 6.0-8.8, 15-50°C. The separation conditions are as follows: stationary phase: pH 4.0 Tris-HCl buffer phase containing 50 mM Tris-HCl and 50 mM KCl; mobile phase A: isooctane containing 50 mM anionic surfactant sodium di(2-ethylhexyl)sulfosuccinate; mobile phase B: 50 mM Tris-HCl buffer containing 500 mM KCl (pH 8.0); In total, 25 mL (23.9 mg) crude enzyme was injected through the injection valve, the enzymatic reaction and sodium dodecylsulfate polyacrylamide gel electrophoresis results imply that the activity of purified α-glucosidase is 6.63-fold higher than that of the crude enzyme. Therefore, countercurrent chromatography coupled with a reverse micelle solvent is capable for protein separation and enrichment. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|