Adaptation of an HPLC system for transient-state enzyme kinetic experiments: pulse-flow method

  • Tsygankov A
  • Motorin Y
  • Wolfson A
 et al. 
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Abstract

The components of an HPLC system can be easily reasembled into an instrument for transient-state enzyme kinetic experiments based on the continuous-flow technique. The scheme for reassembly, operation and data evaluation is described in details for the Pharmacia FPLC system. The working quality of the suggested scheme was tested using a well-studied process of bacterial β-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. © 1990.

Author-supplied keywords

  • Automated assays
  • Enzyme kinetics
  • Mg2+-activation
  • Pulse-flow method
  • β-Galactosidase

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