Fig. 6: Degradation profiles of nPET_GFa/GFc/b/c treated with npFraC_m1/m2.

a,b, HPLC profiles of degradation products released by npFraC_m1 (a) or npFraC_m2 (b) under the experimental conditions: [npFraC_m1/m2], 1.5 μg ml⁻¹; [nPET_GFa/GFc/b/c], 1.1 mg ml⁻¹; reaction volume, 1,000 μl; T, 40 °C; pH, 7.0 (20 mM HEPES buffer); reaction time, 48 h. The reactions were maintained in 2-ml safe-lock Eppendorf polypropylene tubes (ref. 0030 120.094) in a thermoshaker (model Thermomixer comfort, Eppendorf AG) at 1,000 rpm. Aliquots of 100 μl were obtained, the reactions were stopped by adding 900 μl dimethyl sulfoxide (from Merck Life Science) and the degradation products were immediately analysed by HPLC. Datasets were collected with a Varian Star LC workstation 6.41 (Varian). All reactions and analyses were performed in triplicate (n = 3), with a representative chromatogram per enzyme and nPET particle shown. c,d, Concentration of degradation products found to be present in reaction mixtures with npFraC_m1 (c) or npFraC_m2 (d). Quantification was performed for products shown in a and b with unambiguous identification and for which enough material was recovered for HPLC calibration, namely from T to ETETE (Extended Data Fig. 1); purification was performed by semipreparative HPLC, and the molecular weights and structures were determined by mass spectrometry (Supplementary Fig. 15). Values are plotted as the mean of three independent replicates (n = 3) with the reported error ranges and s.d. calculated using the STDEV.S function in Excel 2019 (calculations and raw data are provided in Source Data). The figure was constructed using Excel 2019. Raw data are shown in Supplementary Data 9.