Cyril CHARLIER, Researcher within the Nuclear Magnetic Resonance team at TBI (Toulouse Biotechnology Institute), Toulouse

I have been a CNRS research fellow since November 2019 within the Nuclear Magnetic Resonance team at TBI (Toulouse Biotechnology Institute) in Toulouse. At the TBI, we have access to the 500 and 800 MHz spectrometers of the METATOUL platform, of which only the second is equipped with a cryoprobe.

The TBI is a laboratory at the forefront of fundamental research in biotechnology ranging from gene discovery to process optimization. Scaling up these processes can be done in collaboration with manufacturers. This is particularly the case of the joint PoPLab laboratory located at the TBI and emanating from the collaboration between the TBI and the company CARBIOS. Research within the collaborative laboratory focuses on fundamental aspects of understanding the functioning of enzymes which can not only degrade plastic but also enable its recycling. In this context, we use NMR to dissect the mechanism of enzymes developed to degrade PET (polyethylene terephthalate). Until now our work has focused on the study of the LCC (Leaf and Branch Compost Cutinase) enzyme previously optimized within the laboratory in order to obtain the best enzyme published to date, the quadruple mutant LCC-ICCG.

In order to study this enzyme of approximately 30 kDa and its different variants, we regularly use the 900 MHz of the INFRANALYTICS platform in Lille, and have also recorded the first spectra at 1.2 GHz. The resolution and sensitivity of these very high field spectrometers combined with the competent reception by the engineer in charge give us access to spectra of a quality superior to what can be expected on our spectrometer. The project thus provided valuable information on the different variants used, and made it possible to visualize their structure, dynamics, interactions and catalytic mechanisms from different angles (backbone, methyls and histidine side chains).

Learn more: DOI 10.1016/j.bpd.2022.07.002

Platform used: RMN, IMEC - ISB - UCCS Lille