We have decided to submit 9 new BioBricks this year, which are separated into 2 categories:
- The chassis for our project is based on the Pseudomonas putida bacterium, for its ability to utilize aromatic molecules, mainly focusing on protocatechuate. We have decided to contribute the the iGEM part repository by submitting the genes encoding the enzymes participating in the protocatechuate degradation pathway, in which Protocatechuate, a toxic molecule for most bacteria, is converted to 3-oxoadipate. One of the genes, pcaB, had 4 PstI restriction cut sites and we did not have enough time to introduce silent mutations to the sequence, hence we did not clone and submit it. All 4 other genes were cloned into the pSB1C3 vector and submitted.
- Another part of our project involves engineering of the LC-Cutinase protein. We have chosen the LC-Cutinase protein as a target for rational mutagenesis for its PET degrading activity, and using the PROSS algorithm produced 4 mutants. We have also designed a codon optimized version of the W.T. protein. We have cloned and submitted all 5 LC-Cutinase variants.