• Bioremediation
• enzyme expression
• PET degradation

Bioremediation is the use of microorganism to consume or break down pollutants in the natural environment. This thesis focuses his study on the PET (Polyethylene terephthalate) pollution and its enzymatic degradation due to PETase enzyme activity produce by Ideonella sakaiensis 201-F6. These aerobic bacteria shown to produce two specific enzymes, IsPETase and MHETase, that allow him to use the PET as a carbon source and brake down the hydrocarbon chain.
The research project aimed to study the expression of wild type IsPETase (WT) and a mutant of the WT enzyme, IsPETase W159H-S238F (M) expressed in Escherichia Coli BL21(D3). These two enzymes were subjected to the same treatment, and samples were taken every 24h and 48h for protein analysis and PET degradation rate quantification. Secondly, with a bioinformatics approach, we aimed to suggest a possible novel mutant of the WT IsPETase that would show high degradative capacity and high enzyme expression in bacterial periplasm. This type of study involved the use of different bioinformatics tool, such us: BLAST, CLUSTAL, Pymol and CHARMM-GUI.