7SH6

Crystal structure of a PET hydrolase mutant from Ideonella Sakaiensis

7SH6 の概要

• エントリーDOI: 10.2210/pdb7sh6/pdb
• 分子名称: Poly(ethylene terephthalate) hydrolase, SULFATE ION (3 entities in total)
• 機能のキーワード: pet hydrolase, pet degradation, protein engineering, machine learning, hydrolase
• 由来する生物種: Ideonella sakaiensis
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 30903.54

構造登録者

Kim, W.,Zhang, Y. (登録日: 2021-10-08, 公開日: 2022-04-06, 最終更新日: 2024-10-16)

主引用文献

Lu, H.,Diaz, D.J.,Czarnecki, N.J.,Zhu, C.,Kim, W.,Shroff, R.,Acosta, D.J.,Alexander, B.R.,Cole, H.O.,Zhang, Y.,Lynd, N.A.,Ellington, A.D.,Alper, H.S.
Machine learning-aided engineering of hydrolases for PET depolymerization.
Nature, 604:662-667, 2022

PubMed Abstract: Plastic waste poses an ecological challenge and enzymatic degradation offers one, potentially green and scalable, route for polyesters waste recycling. Poly(ethylene terephthalate) (PET) accounts for 12% of global solid waste, and a circular carbon economy for PET is theoretically attainable through rapid enzymatic depolymerization followed by repolymerization or conversion/valorization into other products. Application of PET hydrolases, however, has been hampered by their lack of robustness to pH and temperature ranges, slow reaction rates and inability to directly use untreated postconsumer plastics. Here, we use a structure-based, machine learning algorithm to engineer a robust and active PET hydrolase. Our mutant and scaffold combination (FAST-PETase: functional, active, stable and tolerant PETase) contains five mutations compared to wild-type PETase (N233K/R224Q/S121E from prediction and D186H/R280A from scaffold) and shows superior PET-hydrolytic activity relative to both wild-type and engineered alternatives between 30 and 50 °C and a range of pH levels. We demonstrate that untreated, postconsumer-PET from 51 different thermoformed products can all be almost completely degraded by FAST-PETase in 1 week. FAST-PETase can also depolymerize untreated, amorphous portions of a commercial water bottle and an entire thermally pretreated water bottle at 50 ºC. Finally, we demonstrate a closed-loop PET recycling process by using FAST-PETase and resynthesizing PET from the recovered monomers. Collectively, our results demonstrate a viable route for enzymatic plastic recycling at the industrial scale.

PubMed: 35478237
DOI: 10.1038/s41586-022-04599-z

実験手法

X-RAY DIFFRACTION (1.44 Å)