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

IF 2.6 4区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Protein Engineering Design & Selection 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。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Protein Engineering Design & Selection 生物-生化与分子生物学

CiteScore

3.30

自引率

4.20%

发文量

审稿时长

6-12 weeks

期刊介绍： Protein Engineering, Design and Selection (PEDS) publishes high-quality research papers and review articles relevant to the engineering, design and selection of proteins for use in biotechnology and therapy, and for understanding the fundamental link between protein sequence, structure, dynamics, function, and evolution.