Fig. 5: The validation of the PET hydrolase with a complex structure based on the iCASE strategy.
a The βTs of PES-H1 under different pressures (1 bar, 100 bar, 500 bar, 1000 bar, 2000 bar, and 4000 bar). n = 3 independent simulations. Error bars represent the means ± SE of βT for PES-H1 under different pressures. b Molecular docking of PES-H1 with 3PET. Hydrogen bonds were depicted by solid blue lines, hydrophobic interactions were illustrated with dashed gray lines, and salt bridges were denoted by dashed yellow lines. c The DSI values of residues of PES-H1. The specific activity (d) and melting temperature (e) of the wild-type PES-H1 and mutants. Reactions were performed in triplicate; Data were presented as mean values ± SD. Data were analyzed by ANOVA and t-test, using the two-tail test. *p < 0.05, **p < 0.01, and ***p < 0.001. The p values of T63H, A64F, S97R, R98L, G99A, Q103L, D107W, S185N, N190L, T63H/A64F, T63H/R98L, A64F/R98L, A64F/S185N, T63H/A64F/R98L, and T63H/A64F/S185N were 1.11 × 10−6, 2.20 × 10−6, 2.08 × 10−5, 1.82 × 10−5, 3.60 × 10−6, 1.16 × 10−7, 1.85 × 10−7, 1.35 × 10−5, 3.93 × 10−1, 1.62 × 10−3, 2.61 × 10−4, 2.66 × 10−3, 8.25 × 10−3, 2.75 × 10−4, and 3.61 × 10−4 in (e), respectively. f Scanning electron microscope images of the treated PET film. PET film was incubated with enzyme in 1 mL of 50 mM glycine-NaOH buffer (pH 9.0) at 70 °C for 4 h. Reactions were performed in triplicate. Source data are provided as a Source Data file.
