title: Application of capillary enzyme micro-reactor in enzyme activity and inhibitors studies of glucose-6-phosphate dehydrogenase.
abstract: In this study, we present an on-line measurement of enzyme activity and inhibition of Glucose-6-phosphate dehydrogenase (G6PDH) enzyme using capillary electrophoresis based immobilized enzyme micro-reactor (CE-based IMER). The IMER was prepared using a two-step protocol based on electrostatic assembly. The micro-reactor exhibited good stability and reproducibility for on-line assay of G6PDH enzyme. Both the activity as well as the inhibition of the G6PDH enzyme by six inhibitors, including three metals (Cu(2+), Pb(2+), Cd(2+)), vancomycin, urea and KMnO4, were investigated using on-line assay of the CE-based IMERs. The enzyme activity and inhibition kinetic constants were measured using the IMERs which were found to be consistent with those using traditional off-line enzyme assays. The kinetic mechanism of each inhibitor was also determined. The present study demonstrates the feasibility of using CE-based IMERs for rapid and efficient on-line assay of G6PDH, an important enzyme in the pentosephosphate pathway of human metabolism.
authors.affiliation: Faculty of Chemistry, Northeast Normal University, Changchun, Jilin 130024, China.
authors.forename: Mohamed Amara
authors.initials: MA
authors.lastname: Camara
authors.affiliation: Faculty of Chemistry, Northeast Normal University, Changchun, Jilin 130024, China.
authors.forename: Miaomiao
authors.initials: M
authors.lastname: Tian
authors.affiliation: Faculty of Chemistry, Northeast Normal University, Changchun, Jilin 130024, China.
authors.forename: Liping
authors.initials: L
authors.lastname: Guo
authors.affiliation: Faculty of Chemistry, Northeast Normal University, Changchun, Jilin 130024, China. Electronic address: yangl330@nenu.edu.cn.
authors.forename: Li
authors.initials: L
authors.lastname: Yang
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D019075:Electrophoresis, Capillary / Q000379:methods*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D019298:Glucose-6-Phosphate / Q000032:analysis / Q000378:metabolism; D005954:Glucosephosphate Dehydrogenase / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D015203:Reproducibility of Results
chemical_list: D004800:Enzymes, Immobilized; D019298:Glucose-6-Phosphate; D005954:Glucosephosphate Dehydrogenase
keywords: Capillary electrophoresis; Enzyme assay; Enzyme micro-reactor; Glucose-6-phosphate dehydrogenase; Inhibition kinetics