Enzymatic Reactions Observed with Zero- and Low-Field Nuclear Magnetic Resonance

Area of Science:

• Biophysical Chemistry
• Analytical Chemistry
• Biochemistry

Background:

• Nuclear Magnetic Resonance (NMR) is a powerful tool for molecular analysis.
• Parahydrogen-induced polarization (PHIP) enhances NMR signal sensitivity.
• Zero- to ultralow-field (ZULF) NMR offers unique advantages over high-field NMR, including signal penetration through conductive materials and reduced line broadening.

Purpose of the Study:

• To demonstrate the capability of ZULF NMR for observing enzyme-catalyzed reactions.
• To investigate the conversion of fumarate to malate and pyruvate to lactate using ZULF NMR.
• To compare zero-field and low-field detection methods for reaction monitoring.

Main Methods:

• Integration of parahydrogen-based hyperpolarization techniques with advanced magnetometry.
• Development of a microfluidic ZULF NMR setup for microliter-scale samples.
• Monitoring of model biological reactions: fumarate to malate and pyruvate to lactate conversion.

Main Results:

• Successful observation of enzyme-catalyzed reactions using ZULF NMR.
• Demonstration of ZULF NMR for monitoring biological processes beyond hydrogenation reactions.
• Comparison of signal detection in zero-field versus low-field NMR regimes.

Conclusions:

• ZULF NMR, combined with hyperpolarization, is a viable technique for monitoring enzyme-catalyzed reactions.
• This approach expands the application of ZULF NMR for general reaction monitoring in biological systems.
• The study highlights the potential of ZULF NMR for sensitive analysis of metabolic processes.