title: Determination of the inhibitory effect of green tea extract on glucose-6-phosphate dehydrogenase based on multilayer capillary enzyme microreactor.
abstract: Natural herbal medicines are an important source of enzyme inhibitors for the discovery of new drugs. A number of natural extracts such as green tea have been used in prevention and treatment of diseases due to their low-cost, low toxicity and good performance. The present study reports an online assay of the activity and inhibition of the green tea extract of the Glucose 6-phosphate dehydrogenase (G6PDH) enzyme using multilayer capillary electrophoresis based immobilized enzyme microreactors (CE-IMERs). The multilayer CE-IMERs were produced with layer-by-layer electrostatic assembly, which can easily enhance the enzyme loading capacity of the microreactor. The activity of the G6PDH enzyme was determined and the enzyme inhibition by the inhibitors from green tea extract was investigated using online assay of the multilayer CE-IMERs. The Michaelis constant (Km ) of the enzyme, the IC50 and Ki values of the inhibitors were achieved and found to agree with those obtained using offline assays. The results show a competitive inhibition of green tea extract on the G6PDH enzyme. The present study provides an efficient and easy-to-operate approach for determining G6PDH enzyme reaction and the inhibition of green tea extract, which may be beneficial in research and the development of natural herbal medicines. Copyright © 2016 John Wiley & Sons, Ltd.
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: Xiaoxia
authors.initials: X
authors.lastname: Liu
authors.affiliation: Faculty of Chemistry, Northeast Normal University, Changchun, Jilin, 130024, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
authors.affiliation: Faculty of Chemistry, Northeast Normal University, Changchun, Jilin, 130024, China.
authors.forename: Yujia
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Faculty of Chemistry, Northeast Normal University, Changchun, Jilin, 130024, China.
authors.forename: Jiqing
authors.initials: J
authors.lastname: Yang
authors.affiliation: Faculty of Chemistry, Northeast Normal University, Changchun, Jilin, 130024, China.
authors.forename: Li
authors.initials: L
authors.lastname: Yang
mesh_terms: D019149:Bioreactors; D019075:Electrophoresis, Capillary / Q000295:instrumentation*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005954:Glucosephosphate Dehydrogenase / Q000037:antagonists & inhibitors*; D010936:Plant Extracts / Q000494:pharmacology*; D013662:Tea / Q000737:chemistry*
chemical_list: D004791:Enzyme Inhibitors; D010936:Plant Extracts; D013662:Tea; D005954:Glucosephosphate Dehydrogenase
keywords: capillary electrophoresis; enzyme microreactor; glucose-6-phosphate dehydrogenase; green tea extract; inhibition kinetics