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

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS ACS Applied Bio Materials Pub Date : 2021-02-15 DOI:10.1093/protein/gzab022
Sebastian Weigert, Andreas Gagsteiger, Teresa Menzel, Birte Höcker
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引用次数: 3

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.

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一个通用的酶促聚对苯二甲酸乙酯降解测定平台。
塑料和微塑料的累积是一个重大的环境挑战。随着高效聚对苯二甲酸乙二醇酯(PET)降解酶的发现,为环境分解和技术回收开辟了新的前景。为了探索自然界中潜在的pet降解酶的巨大多样性,并方便地应用蛋白质工程和定向进化等技术,需要一个快速可靠的检测平台。在这项研究中,我们提出了我们的通用解决方案,将PET涂层应用于标准实验室消耗品,如聚合酶链反应管,96孔和384孔微滴板,产生可调节的PET结晶度。将反应容器与超高性能液相色谱(UHPLC)或荧光读数和额外的酶定量相结合,提供了一系列优势。因此,该平台可以很容易地适应各种需求,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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来源期刊
ACS Applied Bio Materials
ACS Applied Bio Materials Chemistry-Chemistry (all)
CiteScore
9.40
自引率
2.10%
发文量
464
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