title: Adaptation of an HPLC system for transient-state enzyme kinetic experiments: pulse-flow method.
abstract: The components of an HPLC system can be easily reassembled into an instrument for transient-state enzyme kinetic experiments based on the continuous-flow technique. The scheme of reassembly, operation and data evaluation is described in detail for the Pharmacia FPLC system. The working quality of the suggested scheme was tested using a well-studied process of bacterial beta-galactosidase activation by Mg2+ ions. The found parameters were in a good agreement with the literature data. The pulse-flow mode was developed for the delivery of working solutions to reduce the required quantities of enzyme and/or other reaction components. The work presents new approach to HPLC modules as building blocks for sophisticated liquid flow reactors, such as automatic solid phase synthesizers, sequencers and so on.
authors: A. N. Bakh Institute of Biochemistry, Academy of Sciences of the U.S.S.R., Moscow.
authors: Tsygankov AYu
authors: Motorin YuA
authors: A D
authors: AD
authors: Wolfson
authors: D B
authors: DB
authors: Kirpotin
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation*; D004789:Enzyme Activation; D004798:Enzymes / Q000378:metabolism*; D004867:Equipment Design; D007700:Kinetics; D008274:Magnesium / Q000502:physiology; D011673:Pulsatile Flow; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D004798:Enzymes; D001616:beta-Galactosidase; D008274:Magnesium