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.

We interviewed experts, educated the public, and made certain our science would fit what society needs.

The full data and methodology are on our Human Practices page.