Abstract
A new method for the determination of active-enzyme concentration of a glucosidase by using (13)C NMR spectroscopy is reported. The method consists of quantifying the binding between a (13)C-labelled, strong competitive inhibitor, [5-(13)C]-1-azafagomine (1), and the enzyme. The concentration of free inhibitor 1 is measured in a series of binding experiments from the intensity of its NMR signal relative to that of a reference. From a plot of the concentrations of bound vs. free inhibitor 1, the amount of specifically bound 1, that is, the amount of active sites, is determined. From this value, active-enzyme concentration and k(cat) value can be calculated.
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
