Related Experiment Video
Updated: Jun 28, 2026

Multi-enzyme Screening Using a High-throughput Genetic Enzyme Screening System
Published on: August 8, 2016
A bypass trapped-flow analysis system evaluation of enzyme kinetic parameters with a coupled enzyme assay and
1Department of Biochemistry and Anaerobic Microbiology, Virginia Polytechnic Institute and State University, 304 Engel Hall, Blacksburg, VA 24060, USA.
Abstract:
The newly developed flow instrument, bypass trapped-flow analysis system (ByT-FAS), was used to perform enzyme assays and generate data for evaluation of an enzyme's kinetic parameters. The enzyme assay was yeast hexokinase coupled to glucose-6-phosphate dehydrogenase and the production of reduced NADPH was monitored fluorometrically. The kinetic parameters of hexokinase that were evaluated with the system were the apparent K(m)'s of the substrates glucose and ATP, and the K(i) of the competitive inhibitor for ATP, ADP. The ByT-FAS generated data were comparable to other manually derived published values of the kinetic constants for hexokinase and validates the utility of the ByT-FAS system for making absolute enzyme measurements. The semi-automated ByT-FAS system can perform analyses more rapidly than manual assay techniques with less manual manipulation and smaller sample and/or reagent volumes.
Related Concept Videos
Enzyme Kinetics
Scientists typically study enzyme kinetics with a fixed amount of enzyme in the controlled environment of a test tube. When more reactant, or substrate, is...
Fast Reactions
Introduction to Enzyme Kinetics
The experimenter can then plot the initial reaction rate or velocity (Vo) of a given trial against the substrate concentration ([S]) to obtain a graph of the reaction properties. For many enzymatic reactions involving a...

