Time-resolved magnetic resonance to investigate dynamic events in biological systems and biotransformations

Coordinatore di progetto: Università degli Studi di Firenze

Ambiti di ricerca: Chimica, Biochimica e Botanica

Responsabile Scientifico: Elena Babini

Durata: 28/09/2023 - 28/09/2025

Gruppo di ricerca: Enrico Luchinat, Ester Maria Vasini.

Time is a critical dimension in chemistry and biology. Studying the composition of chemical systems as they evolve, including the effects of processing, use, and wear, provides access to the mechanisms underlying the transformations, either wanted or unwanted, as they occur. If monitoring of molecular events can occur in real-time, it can allow for adjusting the reaction conditions to maximize the wanted transformation and minimize the unwanted ones. In biologi-cal systems, which are constantly out of equilibrium, real-time monitoring provides precious insights into the kinetics of the underlying molecular events. Therefore, real-time approaches can radically change the approach to studying chemical and biological processes, paving the way to increased sustainability and a better understanding of living systems. However, selectively observing molecular events which take place within mixtures of high complexity is a chal-lenging task. Commonly-used spectroscopies (UV-VIS, IR) are defeated by the fact that reactants, intermediates, and products have largely superimposable structures, which in turn are reflected in spectra that are difficult to disentan-gle, and the relation between the spectral response and concentrations must be identified (absorption coefficients). On the contrary, NMR is uniquely suited for characterizing the complex responses of these systems. It is intrinsically quantitative, as all the active nuclei provide a signal proportional to their concentration regardless of the chemical environment they are found in, and it is also perfectly selective as even the smallest structural perturbation is re-flected in detectable shifts. Finally, its non-destructive nature makes it ideal for application to biological samples. His-torically, however, NMR has not been included in online reaction monitoring applications because of its intrinsic low sensitivity, which in turn yields long measurement times, and the application to real-time monitoring of living cells is still in its infancy.

In this project, we will develop new acquisition and processing schemes with the aim of reducing the time burden of the NMR experiments while preserving the level of attainable information, thereby pushing the usability of real-time NMR methodologies for real-time monitoring of chemical reactions and biological processes. In particular, we will fo-cus on the development and implementation of blind-source-separation (BSS) methods and fast NMR experiments, and to apply them to a number of biotechnologically-relevant test cases including target-based drug screening in-cell, food processing, and biotransformations. We expect that the methods we propose will lead to a quick maturation of the application for real-time monitoring and that, in the longer run, they might become a standard for time-resolved NMR investigations.