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<p>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. </p>
<p>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.</p>
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