基质(水族馆)
聚对苯二甲酸乙二醇酯
催化作用
组合化学
热稳定性
结晶度
化学
酶动力学
活动站点
立体化学
生物物理学
材料科学
结晶学
酶
有机化学
海洋学
复合材料
生物
地质学
作者
Kun Chen,Yang Hu,Xiaoyan Dong,Yan Sun
标识
DOI:10.1021/acscatal.1c01062
摘要
The accumulation of polyethylene terephthalate (PET) in the environment has brought an enormous threat to the global ecosystem. Although the recently reported PET hydrolase (PETase) displays an efficient decomposition to PET, the low activity and thermostability limit its practical applications. Herein, we introduce a biomodification strategy by fusing a zwitterionic polypeptide (5–30 kDa) consisting of alternating-charged glutamic acid (E) and lysine (K) residues to the C-terminus of PETase and find that increasing the fusion peptide length leads to the improved catalytic performance. The product release in the degradation of highly crystallized PET films (45.2% in crystallinity) by PETase-EK30 is promoted by over 11 times as compared with PETase. The molecular mechanism of the enhanced catalytic performance is investigated via structural analysis, substrate binding, and molecular simulations. Characterizations of the secondary and tertiary structures verify a strengthened structural stability of the EKylated PETases. Synchronous fluorescence spectra indicate a more open substrate-binding pocket after EKylation. MD simulations of enzyme–substrate complexes support that the EKylation induces the exposure of hydrophobic amino acids (W185, I208, and W159) in the substrate-binding pocket and the rotation of the benzene ring of Y87, which promote the substrate binding kinetics. This leads to the enhanced substrate affinity, exactly represented by the increased association constant and the decreased binding free energy. Besides, a shortened catalytic distance is observed from the MD simulations, which might also contribute to the enhanced catalytic activity toward PET degradation. The molecular insights into the enhanced enzyme performance would benefit in extending the application of the EKylation strategy in various enzymes.
科研通智能强力驱动
Strongly Powered by AbleSci AI