9LMT

Crystal structure of variant FAST-ACC-A248E

9LMT の概要

• エントリーDOI: 10.2210/pdb9lmt/pdb
• 分子名称: Poly(ethylene terephthalate) hydrolase (2 entities in total)
• 機能のキーワード: hydrolase, pet hydrolase, pet degradation enzyme
• 由来する生物種: Piscinibacter sakaiensis
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 27392.46

構造登録者

Li, X.,Ning, Z.Y.,Huang, S.Q.,Zeng, C.,Zeng, Z.Y.,Ji, R.,Huang, J.-W.,Chen, C.-C.,Guo, R.-T. (登録日: 2025-01-20, 公開日: 2025-07-23, 最終更新日: 2025-07-30)

主引用文献

Li, X.,Huang, J.W.,Ning, Z.,Huang, S.,Zeng, C.,Zeng, Z.,Ji, R.,Peng, R.,Liu, X.,Min, J.,Chen, C.C.,Guo, R.T.
Combined approaches to enhance the Pichia pastoris-expressed PET hydrolase.
Int.J.Biol.Macromol., 320:145862-145862, 2025

PubMed Abstract: Enzymatic degradation of polyethylene terephthalate (PET) provides a sustainable and promising strategy for the recycling of plastic waste. Herein, we employed site-directed mutagenesis and machine learning methods to further enhance the performance of an efficient mutant of IsPETase, FAST-PETase-N212A, and resulting in seven variants with enhanced activity. We also found that the α3-β5 loop containing the T140D mutation plays a significant role in both type I and type II cutinases. Subsequently, we determined the complex structures of two activity-elevated mutants with the PET analogue mono(2-hydroxyethyl)terephthalic acid, revealing a different binding mode. Finally, to facilitate the industrial application of PET hydrolases, we exploited the industrial strain Pichia pastoris to express the activity-enhanced mutants. Compared with E. coli-produced proteins, these mutants expressed by P. pastoris exhibited higher activity and thermal stability.

PubMed: 40653245
DOI: 10.1016/j.ijbiomac.2025.145862

実験手法

X-RAY DIFFRACTION (1.5 Å)