聚对苯二甲酸乙二醇酯水解酶的结构与功能表征。
Structural and functional characterization of polyethylene terephthalate hydrolase from Ideonella sakaiensis.
机构信息
Department of Biophysics, The Health Science Center, Peking University, Beijing, 100191, China.
Department of Biophysics, The Health Science Center, Peking University, Beijing, 100191, China; Electron Microscopy Analysis Laboratory, The Health Science Center, Peking University, Beijing, 100191, China; Center for Protein Science, Peking University, Beijing, 100871, China.
出版信息
Biochem Biophys Res Commun. 2019 Jan 1;508(1):289-294. doi: 10.1016/j.bbrc.2018.11.148. Epub 2018 Nov 27.
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, IsPETase, and IsPETase mutants can hydrolyze naphthyl esters. IsPETase engineering can direct researchers to use this α/β-hydrolase protein scaffold to design enzymes that can hydrolyze a variety of polyesters.
聚对苯二甲酸乙二醇酯（PET）水解酶来自于解淀粉芽孢杆菌（IsPETase），可以用来降解 PET。为了在工业中使用 IsPETase，我们研究了在自然界中常见的环境和物理化学因素存在的不同条件下，IsPETase 的酶活性。我们观察到盐和甘油增强了酶活性，而清洁剂和有机溶剂降低了酶活性。IsPETase 水解对硝基苯酯（p-NP 酯）而不是萘酯。为了使 IsPETase 成为能够水解萘酯的酶，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、IsPETase 和 IsPETase 突变体可以水解萘酯。IsPETase 工程可以指导研究人员利用这种 α/β-水解酶蛋白支架来设计能够水解各种聚酯的酶。
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