Enzyme kinetics by real-time quantitative NMR (qNMR) spectroscopy with progress curve analysis

Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved..

This review article summarizes how the experimental data obtained using quantitative nuclear magnetic resonance (qNMR) spectroscopy can be combined with progress curve analysis to determine enzyme kinetic parameters. The qNMR approach enables following the enzymatic conversion of the substrate to the product in real-time by a continuous collection of spectra. The Lambert-W function, a closed-form solution to the time-dependent substrate/product kinetics of the rate equation, can estimate the Michaelis-Menten constant (KM.) and the maximum velocity (Vmax) from a single experiment. This article highlights how the qNMR data is well suited for analysis using the Lambert-W function with three different applications. Results from studies on acetylcholinesterase (acetylcholine to acetic acid and choline), β-Galactosidase (lactose to glucose and galactose), and invertase (sucrose to glucose and fructose) are presented. Furthermore, an additional example of how the progress curve analysis is applied to understand the inhibitory role of the artificial sweetener sucralose on sucrose's enzymatic conversion by invertase is discussed. With the wide availability of NMR spectrometers in academia and industries, including bench-top systems with permanent magnets, and the potential to enhance sensitivity using dynamic nuclear polarization in combination with ultrafast methods, the NMR-based enzyme kinetics could be considered a valuable tool for broader applications in the field of enzyme kinetics.

Medienart:

E-Artikel

Erscheinungsjahr:

2022

Erschienen:

2022

Enthalten in:

Zur Gesamtaufnahme - volume:658

Enthalten in:

Analytical biochemistry - 658(2022) vom: 01. Dez., Seite 114919

Sprache:

Englisch

Beteiligte Personen:

Vang, Justin Y [VerfasserIn]
Breceda, Candido [VerfasserIn]
Her, Cheenou [VerfasserIn]
Krishnan, V V [VerfasserIn]

Links:

Volltext

Themen:

β-Galactosidase
30237-26-4
57-50-1
Acetylcholine
Acetylcholinesterase
Beta-Fructofuranosidase
Beta-Galactosidase
Choline
EC 3.1.1.7
EC 3.2.1.23
EC 3.2.1.26
Enzyme kinetics
Fructose
Galactose
Glucose
IY9XDZ35W2
Invertase
J2B2A4N98G
Journal Article
Lactose
N91BDP6H0X
N9YNS0M02X
Nuclear magnetic resonance (NMR)
Progress curve analysis
Review
Sucrose
Sweetening Agents
X2RN3Q8DNE

Anmerkungen:

Date Completed 25.10.2022

Date Revised 30.11.2022

published: Print-Electronic

Citation Status MEDLINE

doi:

10.1016/j.ab.2022.114919

funding:

Förderinstitution / Projekttitel:

PPN (Katalog-ID):

NLM346710952