title: Purification of α-glucosidase from mouse intestine by countercurrent chromatography coupled with a reverse micelle solvent system.
abstract: 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.
authors.affiliation: School of Pharmaceutical Sciences, Southwest University, Chongqing, China.|Department of Clinical Laboratory, Hunan University of Medicine, Hunan, China.
authors.forename: Kai
authors.initials: K
authors.lastname: He
authors.affiliation: School of Pharmaceutical Sciences, Southwest University, Chongqing, China.
authors.forename: Zongyao
authors.initials: Z
authors.lastname: Zou
authors.affiliation: School of Pharmaceutical Sciences, Southwest University, Chongqing, China.
authors.forename: Yinran
authors.initials: Y
authors.lastname: Hu
authors.affiliation: Department of Clinical Laboratory, Hunan University of Medicine, Hunan, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Yang
authors.affiliation: School of Pharmaceutical Sciences, Southwest University, Chongqing, China.|Department of Clinical Laboratory, Hunan University of Medicine, Hunan, China.
authors.forename: Yubo
authors.initials: Y
authors.lastname: Xiao
authors.affiliation: Department of Clinical Laboratory, Hunan University of Medicine, Hunan, China.
authors.forename: Pincao
authors.initials: P
authors.lastname: Gao
authors.affiliation: School of Pharmaceutical Sciences, Southwest University, Chongqing, China.
authors.forename: Xuegang
authors.initials: X
authors.lastname: Li
authors.affiliation: School of Life Sciences, Southwest University, Chongqing, China.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Ye
mesh_terms: D000818:Animals; D002845:Chromatography / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D003377:Countercurrent Distribution / Q000379:methods; D004573:Electrolytes; D007422:Intestines / Q000201:enzymology*; D008027:Light; D051379:Mice; D008823:Micelles; D012031:Refractometry; D012492:Salts / Q000737:chemistry; D012542:Scattering, Radiation; D012997:Solvents / Q000737:chemistry*; D000520:alpha-Glucosidases / Q000737:chemistry*
chemical_list: D004573:Electrolytes; D008823:Micelles; D012492:Salts; D012997:Solvents; D000520:alpha-Glucosidases
keywords: High-speed countercurrent chromatography; Protein purification; Reverse micelles; alpha-Glucosidase