title: Use of capillary electrophoresis and indirect detection to quantitate in-capillary enzyme-catalyzed microreactions.
abstract: The use of capillary electrophoresis and indirect detection to quantify reaction products of in-capillary enzyme-catalyzed microreactions is described. Migrating in a capillary under conditions of electrophoresis, plugs of enzyme and substrate are injected and allowed to react. Capillary electrophoresis is subsequently used to measure the extent of reaction. This technique is demonstrated using two model systems: the conversion of fructose-1,6-bisphosphate to dihydroxyacetone phosphate and glyceraldehyde-3-phosphate by fructose-biphosphate aldolase (ALD, EC 4.1.2.13), and the conversion of fructose-1,6-bisphosphate to fructose-6-phosphate by fructose-1,6-bisphospatase (FBPase, EC 3.1.3.11). These procedures expand the use of the capillary as a microreactor and offer a new approach to analyzing enzyme-mediated reactions.
authors.affiliation: Department of Chemistry and Biochemistry, California State University, Los Angeles 90032-8002, USA.
authors.forename: Y
authors.initials: Y
authors.lastname: Zhang
authors.forename: M R
authors.initials: MR
authors.lastname: el-Maghrabi
authors.forename: F A
authors.initials: FA
authors.lastname: Gomez
mesh_terms: D004099:Dihydroxyacetone Phosphate / Q000737:chemistry; D019075:Electrophoresis, Capillary; D006597:Fructose-Bisphosphatase / Q000737:chemistry; D005634:Fructose-Bisphosphate Aldolase / Q000737:chemistry; D005635:Fructosediphosphates / Q000737:chemistry; D005636:Fructosephosphates / Q000737:chemistry; D005986:Glyceraldehyde 3-Phosphate / Q000737:chemistry
chemical_list: D005635:Fructosediphosphates; D005636:Fructosephosphates; D005986:Glyceraldehyde 3-Phosphate; D004099:Dihydroxyacetone Phosphate; C027618:fructose-6-phosphate; D006597:Fructose-Bisphosphatase; D005634:Fructose-Bisphosphate Aldolase; C029063:fructose-1,6-diphosphate