Ingegnerizzazione del citocromo P450 4F11 per applicazioni biotecnologiche: Produzione di Bioplastiche

Omega hydroxyfatty acids are excellent monomers for synthesizing a unique family of polyethylene-like biobased plastics. However, ω-hydroxyfatty acids are difficult and expensive to prepare by traditional organic synthesis, precluding their use in commodity materials. Here we report the engineering of the human cytochrome P450 4F11 to study the ability to perform ω-hydroxylation of medium/long chain fatty acids. A purification system was developed for human P450 4F11, producing ~8 mg P450 from a 1-liter culture of Escherichia coli. Two fatty acids¿palmitic and oleic¿were identified in human liver and verified as substrates of P450 4F11. The products were characterized as ω-hydroxylated fatty acids by Fluorescence measurements using an experimental probe kindly donated by Cage Chemicals s.r.l. Further our results show that the P450 4F11 has around the 40% of the heam cofactor covalently bound to the protein. According to the literature structural homology model of the protein reveal a strictly conserved glutamic acid residue in the I helix positioned with a useful orientation for the bound with the haem.