Our new Composite Part is a significant contribution.
Our composite part fuses the ICCG and TfCa enzyme domains, neither previously paired with dockerin/cohesin scaffolding of this kind, onto a shared cohesin scaffold via dockerin linkers, so that PET and MHET hydrolysis can occur in a coordinated sequence rather than as two separate, uncoordinated reactions. This scaffolding approach is, to our knowledge, new to the iGEM registry, and future teams working on multi-enzyme plastic degradation could adapt the same scaffold to co-localize other enzyme pairs.
The full construct sequences and design rationale are on our Composite Part page.
We characterized our constructs using SDS-PAGE, Western blot, and quantitative BSA assay for protein identity and purity, and used both qualitative and quantitative assays, including a standard TPA curve, to measure PETase activity. In at least two cases, we believe these part-level measurements are precise and well-controlled enough that other teams could build directly on them rather than re-measuring from scratch.
The full data and methodology are on our Measurement page.