一种聚对苯二甲酸乙二醇酯(PET)水解新酶的挖掘与酶学性质分析
作者:
作者单位:

1.天津科技大学 生物工程学院,天津;2.天津中医药大学 研究生院,天津;3.中国科学院天津工业生物技术研究所,低碳合成工程生物学全国重点实验室,天津

作者简介:

赵伟:PET水解酶的挖掘、性能测试及文章撰写;毕海珍:PET水解酶的克隆表达;Yew Maxine:PET水解酶的活性测试;翟欢欢:部分数据分析及文章修改;张翠英:实验设计及文章修改;朱蕾蕾:整体实验设计及文章修改。

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基金项目:

国家重点研发计划(2023YFC3905000)


Discovery and characterization of a new enzyme for hydrolyzing polyethylene terephthalate (PET)
Author:
Affiliation:

1.School of Biological Engineering, Tianjin University of Science & Technology, Tianjin, China;2.Graduate School, Tianjin University of Traditional Chinese Medicine, Tianjin, China;3.State Key Laboratory of Engineering Biology for Low-carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China

Fund Project:

This work was supported by the National Key Research and Development Program of China (2023YFC3905000).

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    摘要:

    目的 聚对苯二甲酸乙二醇酯(polyethylene terephthalate, PET)的生物酶法解聚是通过PET水解酶催化PET水解,得到终产物对苯二甲酸和乙二醇。通过基因挖掘获得新的PET水解酶,可为废弃PET的生物酶法解聚提供可用于分子改造的蛋白。方法 通过基因挖掘的方法发现来源于Alkalilimnicola ehrlichii的未知功能蛋白(命名为AePETase)具有水解无定形PET膜的活性,并对AePETase进行异源表达、纯化和酶学性质研究。结果 确定了AePETase的Tm值为(53.0±0.7) ℃,最适反应温度为45 ℃,最适缓冲液体系为Na2HPO4-NaH2PO4 (100 mmol/L, pH 8.0),较合适的酶浓度为1.0 μmol/L以内。在45 ℃条件下,无定形PET膜的水解产物量为已报道的IsPETase的13.4倍(72 h)。结论 AePETase对PET具有较好的水解活性,是一种具有较大潜力的PET水解新酶。

    Abstract:

    Objective The enzymatic depolymerization of polyethylene terephthalate (PET) involves the hydrolysis of PET catalyzed by PET hydrolases, leading to the production of terephthalic acid and ethylene glycol. In this study, we identified a new PET hydrolase through gene mining, with the goal of providing enzymes for efficient depolymerization of waste PET.Methods A previously uncharacterized functional enzyme, named as AePETase, was identified through gene mining. This enzyme demonstrated the ability to hydrolyze amorphous PET films. Subsequently, heterologous expression, purification, and enzymatic characterization were performed for AePETase.Results AePETase showed a Tm value of (53.0±0.7) ℃, and an optimal reaction temperature of 45 ℃. The optimal buffer was Na2HPO4-NaH2PO4 (100 mmol/L, pH 8.0), and the optimal enzyme concentration was 1 μmol/L. After 72 h, the hydrolysis of amorphous PET films by AePETase resulted in 13.4-fold more products than that by the reported IsPETase.Conclusion AePETase demonstrates significant hydrolytic activity on PET and represents a promising new enzyme for hydrolyzing PET.

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赵伟,毕海珍,Yew Maxine,翟欢欢,张翠英,朱蕾蕾. 一种聚对苯二甲酸乙二醇酯(PET)水解新酶的挖掘与酶学性质分析[J]. 微生物学报, 2025, 65(7): 3126-3135

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  • 收稿日期:2025-01-06
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  • 在线发布日期: 2025-07-04
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