Light-NMR spectroscopy: tools and applications

Lead Research Organisation: University of Manchester
Department Name: Chemistry

Abstract

Light is a form of electromagnetic energy that surrounds and affects matter around us: a myriad of chemical reactions and life processes depend on light. Light is very easy to produce and control, and therefore it can be an ideal trigger that connects our macroscopic world with nanoscale or molecular objects, to modulate material properties, chemical reactions, molecular switches, nano-devices, gene expression, targeted drug release, and many other physical processes. How light modulates matter, and how it can be used to control it, is one of the most important questions in science. New and more powerful analytical approaches are needed to reveal and closely monitor the effects of light on complex chemical systems, for example, photoenzymes emerging from chemical and synthetic biology fields, to explore and optimise the mechanisms involved. NMR spectroscopy is one of the most powerful and universal techniques in chemistry, physics and life sciences to study the properties and structure of matter. NMR spectral parameters are sensitive to chemical and 3D structure of molecules present in the samples, as well as their local environment. Therefore NMR is routinely used for confirming the identity of molecular structures, quality and purity analysis, chemical reaction monitoring, as well as monitoring the changes in conformation and molecular dynamics. The project aims at developing tools for combining illumination with NMR experiments, so that the whole power of NMR can be applied to studies of photochemical and photoenzymatic systems.
 
Description The aim of the project was to develop tools for combining illumination with NMR experiments, so that samples could be effectively illuminated in situ inside the NMR spectrometer. As a part of the project, we have created and tested new multi-channel illumination equipment for NMR samples, which would allow light of different colours (or combinations thereof) to be applied with precise timing, synchronized with NMR pulse programs. We named our equipment "NMRTorch" because it resembles a torch or a flashlight that shines light directly into the NMR sample. We created and tested a four-channel light driver module, as well as a number of auxiliary devices based on microcontrollers, which provided an additional level of control both manually by the operator, and automatically via spectrometer pulse programs and/or experimental queuing systems. Such equipment enabled the automation of experiments with very diverse possible illuminating regimes, including switching light on and off, and toggling between continuous and pulsed illumination using different colours, as well as adjusting light intensity using pulse width modulation. These controls were incorporated into a wide range of NMR experiments (pulse sequence programs), both 1D and 2D. As a part of this project, we also created and tested a number of multi-colour light sources (light heads) with different combinations of light-emitting diodes (LEDs) in both visible range and UV, allowing for the study of a wide range of photo-responsive systems. Light heads housing two, three, and four LEDs were designed and successfully tested, in the range from UV (280nm) to near-infrared. Moreover, we created versions of the NMRTorch light heads that are suitable for benchtop NMR spectrometers and tested them successfully, proving that the approach works across different spectrometers and magnetic fields (800 MHz, 500 MHz, and 60 MHz). We tested NMRTorch on a number of photoreactive molecules, such as photoswitches, photoenzymes, and other photo-responsive systems. For example, it allowed us to characterize the conversion kinetics in systems such as amino-azobenzene, which shifts cis-trans equilibrium depending on light colour, and study the photodegradation of quinine molecules. In collaboration with other groups, we tested NMRTorch for monitoring kinetics of chemical reactions catalysed by photoenzyme created by directed evolution, and kinetics of a family of azobenzenes where chemical substitutions have modulated the rates of photoswitching and thermal relaxation, as well as photostationary states. We demonstrated that the intensity of light delivered inside the NMR sample can be measured using photochemical reaction with known quantum yield; moreover, we demonstrated that use of additional illumination in UV range (at 280 nm) allows to conveniently reset the starting concentration of a particular isoform, so the concentration series needed for the actinometry experiment can be conducted in automated manner using just one sample. Importantly, the intensity of light delivered to the NMR sample can be very high: for 365 nm substantial quinine degradation can be achieved in less than 2 hours, meaning that photostability testing to pharmaceutical standards can be performed inside NMR spectrometer, with live monitoring of the process. In situ illumination by the 365 nm LED NMRtorch enabled effective and controlled release of photocaged ATP with simultaneous monitoring of subsequent enzymatic reactions by NMR spectroscopy. Overall, we demonstrated applicability of NMRTorch approach for diverse photo-reactive systems.
Exploitation Route The novel NMRTorch approach developed and tested in this project may be used by both academia and industry to study photo-responsive systems in situ and in operando using solution-state NMR spectroscopy. Furthermore, the NMRTorch devices and consumables (patent pending) in principle can be produced commercially. Currently, the University of Manchester offers a license for their production.
Sectors Agriculture, Food and Drink,Chemicals,Education,Electronics,Energy,Environment,Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology
URL https://doi.org/10.1038/s42004-022-00704-5
 
Description The results of this project represent a breakthrough by having solved a fundamental research challenge: how to apply NMR spectroscopy universally to complex photo-responsive systems. It may take time for these results to make a wider impact. It is too early to discuss non-academic impacts since the first papers describing the results were published only recently. Licenses for the technology at the moment are available for commercialization by any interested parties.
First Year Of Impact 2022
Sector Chemicals,Energy,Environment,Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology
 
Description UKRI (EPSRC) Impact Acceleration Account: Relationship Development Scheme, ref 388
Amount £12,542 (GBP)
Funding ID 388 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 01/2023 
End 04/2023
 
Title A device has been created for illuminating NMR samples in situ. 
Description We have designed, made and tested prototypes of devices (power supplies and lightheads) for multiwavelength illumination NMR samples in situ in NMR spectrometer. The patent application PCT/GB2021/051254 describing this device has been published in December 2021. The first research papers describing the approach in more detail also have been published. The device can be used locally by collaborating research groups. 
Type Of Material Improvements to research infrastructure 
Year Produced 2021 
Provided To Others? Yes  
Impact No specific impacts yet to report, apart from first research papers published and in preparation. 
URL https://www.nature.com/articles/s42004-022-00704-5
 
Description Collaboration with AstraZeneca 
Organisation AstraZeneca
Department Research and Development AstraZeneca
Country United Kingdom 
Sector Private 
PI Contribution We started a collaboration with AstraZeneca, which got subsequently funded by UKRI (EPSRC) Impact Acceleration Account (IAA): Relationship Development Scheme, for the project "Demonstrating the applicability of light-coupled NMR spectroscopy for photo-stability studies of biopharmaceutical formulations". This collaboration is covered by an agreement signed by both sides earlier in 2022.
Collaborator Contribution The Partners on this collaboration, AstraZeneca, have supplied materials required for the experiments, as well as their staff time. Additionally, looking at the emerging NMR results, recently they planned to run a series of complimentary experiments in AstraZeneca so that NMR results can be benchmarked against industry-standard approaches.
Impact No outputs/outcomes yet.
Start Year 2022
 
Title ILLUMINATION INSERT FOR AN NMR SPECTROMETER 
Description An illumination insert for an NMR spectrometer, the illumination insert being shaped to receive a sample and comprising a light guide portion for guiding light from a light source, and a diffuser portion for diffusing light received from the light guide portion towards a sample received in the illumination insert. 
IP Reference WO2021250372 
Protection Patent application published
Year Protection Granted 2021
Licensed No
Impact No impacts yet.
 
Description Chemistry Community @ Nature "Behind the Paper" blog post. 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact "Behind the paper" community blog article "Shining a light on chemistry with the NMRtorch" explaining, mostly to a lay audience, the implications of the research topic in general, and of the paper just published in Nature's Chemistry Communications.
Year(s) Of Engagement Activity 2022
URL https://chemistrycommunity.nature.com/posts/shining-a-light-on-chemistry-with-the-nmrtorch
 
Description Chemistry World Webinar 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Royal Society of Chemistry, Chemistry World webinar "Shine a light on your chemistry with in-situ photo-NMR"
Year(s) Of Engagement Activity 2022
URL https://www.chemistryworld.com/webinars/shine-a-light-on-your-chemistry-with-in-situ-photo-nmr/40151...
 
Description Press release by the University of Manchester 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact Press-release issued by the University of Manchester Press Office outlining the project started which has been just funded by EPSRC, before the actual project begun. The aim of this press-release is to raise the awareness of the community about the type of novel groundbreaking research that we are starting to do.
Year(s) Of Engagement Activity 2020
URL https://www.manchester.ac.uk/discover/news/new-horizons-for-research-through-new-adventurous-researc...