Real-Time Monitoring of Hydrolysis Reactions of Pyrophosphates with Dissolution Dynamic Nuclear Polarization

Area of Science:

• Nuclear Magnetic Resonance Spectroscopy
• Biochemistry
• Chemical Dynamics

Background:

• Dissolution dynamic nuclear polarization (d-DNP) is widely used for real-time reaction monitoring, primarily with carbon-13 and nitrogen-15 spins.
• Phosphorus-31 (31P) applications in d-DNP are limited to pH imaging and crystallization due to short relaxation times.

Purpose of the Study:

• To demonstrate the feasibility of observing enzyme reactions using d-DNP with phosphorus-31 spins.
• To overcome the limitations of short relaxation times for 31P in d-DNP applications.

Main Methods:

• Utilized bullet-DNP to obtain hyperpolarized 31P spins in pyrophosphate, requiring less dilution of solid samples.
• Applied d-DNP for real-time monitoring of pyrophosphate hydrolysis by inorganic pyrophosphatase.

Main Results:

• Successfully achieved real-time monitoring of the pyrophosphate hydrolysis reaction catalyzed by inorganic pyrophosphatase.
• Determined the reaction rate of pyrophosphate hydrolysis at physiological pH.
• Demonstrated the potential for observing enzyme reactions with 31P using d-DNP.

Conclusions:

• Dynamic nuclear polarization with phosphorus-31 can be effectively used to study enzyme reactions.
• This technique opens new avenues for applications in medicine, agriculture, and quantum life sciences.
• Overcoming short relaxation times in 31P enhances the utility of d-DNP in chemical and biological studies.