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一种用于酶法降解聚对苯二甲酸乙二酯的多功能分析平台。

A versatile assay platform for enzymatic poly(ethylene-terephthalate) degradation.

机构信息

Department of Biochemistry, University of Bayreuth, Universitätsstraße 30, 95447 Bayreuth, Germany.

Department of Polymer Engineering, University of Bayreuth, Universitätsstraße 30, 95447 Bayreuth, Germany.

出版信息

Protein Eng Des Sel. 2021 Feb 15;34. doi: 10.1093/protein/gzab022.

Abstract

Accumulation of plastic and subsequent microplastic is a major environmental challenge. With the discovery of potent polyethylene terephthalate (PET)-degrading enzymes, a new perspective arose for environmental decomposition as well as technical recycling. To explore the enormous diversity of potential PET-degrading enzymes in nature and also to conveniently employ techniques like protein engineering and directed evolution, a fast and reliable assay platform is needed. In this study we present our versatile solution applying a PET coating on standard lab consumables such as polymerase chain reaction tubes, 96- and 384-well microtiter plates, yielding an adjustable crystallinity of the PET. Combining the reaction vessels with either ultra-high performance liquid chromatography (UHPLC) or fluorometric readout and additional enzyme quantification offers a range of advantages. Thereby, the platform can easily be adapted to diverse needs from detailed analysis with high precision to high-throughput (HT) applications including crude lysate analysis.

摘要

塑料的积累和随后的微塑料是一个主要的环境挑战。随着发现有效的聚对苯二甲酸乙二醇酯(PET)降解酶,为环境分解以及技术回收提供了新的视角。为了探索自然界中潜在的 PET 降解酶的巨大多样性,并且方便地采用蛋白质工程和定向进化等技术,需要一个快速而可靠的测定平台。在这项研究中,我们提出了一种多功能的解决方案,即将 PET 涂层应用于标准实验室耗材上,如聚合酶链反应管、96 孔和 384 孔微量滴定板,从而使 PET 的结晶度具有可调节性。将反应容器与超高效液相色谱(UHPLC)或荧光检测相结合,并进行额外的酶定量分析,提供了一系列的优势。因此,该平台可以轻松适应各种需求,从高精度的详细分析到高通量(HT)应用,包括粗裂解物分析。

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