Cells were pelleted by centrifugation at 4000 xg for 10 min at 4 ºC and resuspended in Buffer W (100 mM Tris (pH 8.0), 150 mM NaCl) before homogenization using Emulsiflex C5. The lysate was cleared by centrifugation at 20000 xg for 30 minutes and passed through a Strep-Tactin (IBA) gravity column following the producer's protocol for purifying Strep-tag II fusion proteins. The purified DuraPETase was buffer exchanged into carbonate buffer (27 mM Na2CO3, 90 mM NaHCO3 (pH 8.5), 100 mM NaCl) using PD-10 desalting columns (GE Healthcare) and stored at 4 °C for up to 4 days.