The amino acid sequences for the enzymes are shown in Figure . The DuraPETase gene, with a C-terminal Strep-tag, was obtained as a synthetic gene (ThermoFischer) before cloning into the E. coli expression vector pHAT5 with restriction enzymes (ThermoFischer FastDigest) NcoI and XhoI. For the expression and purification of DuraPETase (Figure ), single colonies from E. coli OverExpress TM C41 (DE3) (Lucigen) transformations were grown in MDAG-135 non-inducing media supplemented with 50 μg mL -1 carbenicillin (37 °C; 200 rpm shaking; 24 h) as a starter culture. ZYM-5052 media was inoculated with the starter culture (1:100 v/v inoculation ratio) and incubated at 20 °C with 300 rpm shaking until growth saturation was reached for overexpression.