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Bioelectrochemical approach for characterisation of Nitrogenase-like Enzyme Dark-Operative Protochlorophyllide Reductase (DPOR)

Author Giada BEDENDI
Director of thesis Dr. Ross Dean Milton
Co-director of thesis
Summary of thesis

My phD research project concerns the study of an enzyme, called Dark-Operative Protochlorophyllide Reductase (DPOR), which takes part in chlorophyll photosynthesis. The DPOR enzyme belongs to the family of redox proteins, precious for all life cycles. One of the most known enzyme of this family is Nitrogenase, which in nature is the only enzyme known to date capable of synthesizing ammonia, thus representing a valid candidate for the bioproduction of ammonia.

Despite decades of research dedicated to finding alternative solutions and studying the nitrogenase enzyme, we have not yet been able to understand its reaction mechanism.

The study of the DPOR enzyme, which shares the same structural characteristics with Nitrogenase, could be the key of understanding its mechanism and taking one more step towards a sustainable future.

DPOR is not a widely studied enzyme and the topic is considered quite challenging. During my phD, the following points were optimized: the production of DPOR, assays to verify its enzymatic activity and it was finally characterized spectroscopically and electrochemically. In particular, the characterization of the enzyme is performed through spectroelectrochemistry: a state-of-the-art technique that allows the spectroscopic and electrochemical properties of the enzyme to be analyzed simultaneously and in situ. Through the spectroelectrochemistry it is possible to know the reduction potentials of the redox cofactors present in the subunits of the enzyme, which are essential for understanding the reaction mechanism.

The ultimate goal of the project is to know the properties and mechanism of the DPOR enzyme using a multidisciplinary approach that provides for a good combination of chemistry, biology and physics.

The DPOR enzyme is still poorly studied and this project will bring to light new data and it will also provide the knowledge for an innovative approach for the reduction and recycling of greenhouse gas emissions.

Status beginning
Administrative delay for the defence 2024