A versatile assay platform for enzymatic poly(ethylene-terephthalate) degradation. (24th August 2021)

• Record Type: Journal Article
• Authors: Weigert, Sebastian; Gagsteiger, Andreas; Menzel, Teresa; Höcker, Birte
• Is Part Of: Protein engineering, design & selection. Volume 34:(2021)
• Publication Date: 2021-08-24
• Subjects: assay platform -- PET-degrading enzymes -- PETase -- screening; Protein engineering -- Periodicals; 660.63
• DOI: 10.1093/protein/gzab022
• Languages: English
• ISSNs: 1741-0126

Abstract

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.