About This Project

Polyethylene terephthalate (PET), used in bottles, packaging, and polyester clothing, is difficult to recycle. This project asks whether RT1, a computer-designed enzyme, can break down PET at room temperature. Simulations suggest RT1 holds the shape needed for activity. We hypothesize that it will produce measurable PET breakdown products. Funds will support making and purifying RT1 and testing it on PET. The results will guide the next design cycle.

Ask the Scientists

What is the context of this research?

Polyethylene terephthalate (PET) is used in bottles, food containers, and polyester textiles. Its durability makes discarded PET difficult to break down. The discovery of a bacterium that uses enzymes to degrade PET showed that biological recycling is possible. Researchers have since developed faster, more robust enzymes, but their performance still depends on temperature, pH, and the form of the plastic. RT1 is a computer-designed PETase—a PET-degrading enzyme. Simulations suggest its active site, where breakdown begins, may hold a useful shape at room temperature, but simulations cannot prove activity. We hypothesize that purified RT1 will release measurable PET breakdown products at room temperature. Laboratory testing is needed to determine whether this prediction is correct.

What is the significance of this project?

Computational enzyme design is moving fast, but predictions only matter if they survive contact with the lab. This project is important because it tests whether an AI-designed PETase candidate can become a real, measurable enzyme rather than just a promising model. If RT1 shows activity, it would be an early step toward lower-energy plastic recycling approaches. If it does not, the result is still valuable because it tells us what failed: expression, folding, stability, or catalytic activity. Either outcome creates useful data for the next design cycle.

What are the goals of the project?

The goal is to run the smallest complete experiment that can answer whether RT1 is worth developing further. First, we will synthesize and clone the RT1 gene. Second, we will express the protein in E. coli and purify it using a His-tag. Third, we will expose PET samples to RT1 under controlled conditions. Finally, we will use HPLC analysis to look for PET breakdown products such as TPA or MHET. The outcome will tell us whether RT1 can be produced and whether it shows measurable PET-degrading activity.