Structural and functional characterization of polyethylene terephthalate hydrolase from Ideonella sakaiensis
Structural and functional characterization of polyethylene terephthalate hydrolase from Ideonella sakaiensis
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坂井 Ideonella sakaiensis 聚对苯二甲酸乙二醇酯水解酶的结构和功能表征
DOI：
10.1016/j.bbrc.2018.11.148
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发表时间：
2019-01-01
影响因子：
3.1
通讯作者：
Yin, Chang-Cheng
中科院分区：
文献类型：
--
作者：
Liu, Congcong;Shi, Chao;Yin, Chang-Cheng
日本产的聚对苯二甲酸乙二醇酯(PET)水解酶(IsPETase)可用于降解PET。为了将IsPETase用于工业生产，我们研究了IsPETase在不同环境和自然界中常见的物理化学因素下的酶活性。我们观察到盐和甘油提高了酶的活性，而洗涤剂和有机溶剂降低了酶的活性。IsPETase水解对硝基苯酯(p-NP)酯而不是萘酯。为了使IsPETase成为一种能够水解萘酯的酶，根据晶体结构提供的结构信息进行了定点突变。我们发现，IsPETase(S93M)、ISPETase(W159F)和IsPETase(N241F)突变株具有水解萘酯的能力。IsPETase工程可以指导研究人员使用这种α/β水解酶蛋白支架来设计可以水解多种聚酯的酶。(C)2018 Elsevier Inc.保留所有权利。
Polyethylene terephthalate (PET) hydrolase from Ideonella sakaiensis (IsPETase) can be used to degrade PET. In order to use IsPETase in industry, we studied the enzymatic activity of IsPETase in different conditions containing environmental and physicochemical factors commonly found in nature. We observed that salts and glycerol enhanced the enzymatic activity, while detergents and organic solvents reduced the enzymatic activity. IsPETase hydrolyzed p-nitrophenyl (p-NP) esters instead of naphthyl esters. To make IsPETase an enzyme capable of hydrolyzing naphthyl esters, site-directed mutagenesis was carried out based on the structural information provided by the crystal structure. We found that the IsPETase(s93m), ISPETase(w159F), and IsPETase(N241F) mutants can hydrolyze naphthyl esters. IsPETase engineering can direct researchers to use this alpha./beta-hydrolase protein scaffold to design enzymes that can hydrolyze a variety of polyesters. (C) 2018 Elsevier Inc. All rights reserved.