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Updated: Jun 13, 2026

Dissolution Dynamic Nuclear Polarization Instrumentation for Real-time Enzymatic Reaction Rate Measurements by NMR
Published on: February 23, 2016
A direct NMR method for the measurement of competitive kinetic isotope effects
Jefferson Chan1, Andrew R Lewis, Michel Gilbert
1Department of Chemistry, Simon Fraser University, Burnaby, British Columbia, Canada.
Abstract:
We present a technique that uses (13)C NMR spectroscopy to measure kinetic isotope effects on the second-order rate constant (k(cat)/K(m)) for enzyme-catalyzed reactions. Using only milligram quantities of isotopically labeled substrates, precise competitive KIEs can be determined while following the ongoing reaction directly in a NMR spectrometer. Our results for the Vibrio cholerae sialidase-catalyzed hydrolysis of natural substrate analogs support a concerted enzymatic transition state for these reactions.
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