Workflow for characterization of enzymes under different reaction conditions

An experimental workflow to provide detailed information of the molecular mechanisms of enzymes is described. This workflow will help in the application of enzymes in technical processes by providing crucial parameters needed to plan, model and implement biocatalytic processes more efficiently. These parameters are homogeneity of the enzyme sample (HES), kinetic and thermodynamic parameters of enzyme kinetics and binding of reactants to enzymes. The techniques used to measure these properties are dynamic light scattering (DLS), UV-Vis spectrophotometry and isothermal titration calorimetry (ITC) respectively. The workflow is standardized by the use of SOPs and python-scripted data analysis. We have used the NADPH-dependent alcohol dehydrogenase Gre2p as a challenging enzyme to demonstrate the power of this workflow. Our work highlights the utility for combined binding and kinetic studies for such complex multi-substrate reactions and the importance of sample quality control during experiments.

Help
help Creators and Submitter
Creators
Submitter
Activity

Views: 109

Created: 21st Apr 2021 at 10:06

Last updated: 5th May 2021 at 14:02

Related items

Powered by
(v.1.11.0)
Copyright © 2008 - 2021 The University of Manchester and HITS gGmbH

By continuing to use this site you agree to the use of cookies