Department of Biochemistry, University of Bayreuth, Universitätsstraße 30, 95447 Bayreuth, Germany.
Department of Polymer Engineering, University of Bayreuth, Universitätsstraße 30, 95447 Bayreuth, Germany.
Protein Eng Des Sel. 2021 Feb 15;34. doi: 10.1093/protein/gzab022.
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.
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。