Extended Data Fig. 5: Comparison of HotPETase activity with different substrates at 40 °C and 60 °C under library screening conditions and optimised reaction conditions.
From: Directed evolution of an efficient and thermostable PET depolymerase
Bar chart showing the mean total concentration of released MHET and TPA accumulated over 3 h (light pink) and 24 h (dark pink) at either 40 °C or 60 °C in reactions using HotPETase with different PET substrates (crystalline PET powder (cryPET), milled bottle-grade PET (bgPET), PET/PE composite film lid (PET/PE), 0.4% total substrate loading (4 g L−1)). Reactions were performed using either library hit screening conditions (A): 0.29 mg g−1 enzyme loading (0.04 μM), pH 9.2, 50 mM Gly-OH, 4% BugBuster or optimised conditions (B): 3.62 mg g−1 enzyme loading (0.5 μM), pH 9.7, 50 mM Gly-OH buffer, 4% BugBuster. Reactions were carried out in triplicate; error bars represent the s.d. of the replicate measurements; each replicate measurement is represented as a black circle.
