Cellular and Molecular Biology

Brief description
The project aims at the functional and structural characterization of a cytochrome P450 obtained from the cyanobacterium Stanieria cyanosphaera. 

Based on the structural alignment, we found that this enzyme belongs to the group of P450 peroxygenases, which accept electrons from H₂O₂ to perform their catalytic activity. The main objective of the project is to obtain pure protein in order to (i) find the optimal condition for crystallization (ii) study the thermal stability and the resistance to H₂O₂ (iii) determine the spectroscopical features of the enzyme, (iv) study the enzymatic activity towards its natural substrates (fatty acids), (v) characterize the oxidized products using GC-MS. 

Knowledge, skills and competences to be acquired by the end of the traineeship (expected Learning Outcomes):
The student will acquire competences in 1) protein expression using E. coli, 2) protein purification, 3) protein characterization using different techniques including SDS-PAGE, analytical gel filtration, spectrophotometer, Differential Scanning Calorimetry, crystallography. The student will also study the enzymatic activity and will acquire competences in Gas Chromatography and Mass Spectrometry.