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生物传感器与化学酶一锅法级联应用于检测和转化活细胞中聚对苯二甲酸乙二酯衍生的对苯二甲酸。

Biosensor and chemo-enzymatic one-pot cascade applications to detect and transform PET-derived terephthalic acid in living cells.

作者信息

Bayer Thomas, Pfaff Lara, Branson Yannick, Becker Aileen, Wu Shuke, Bornscheuer Uwe T, Wei Ren

机构信息

Institute of Biochemistry, Department of Biotechnology & Enzyme Catalysis, University of Greifswald, Felix-Hausdorff-Straße 4, 17487 Greifswald, Germany.

Institute of Molecular Biotechnology, TU Graz, Petersgasse 14, 8010 Graz, Austria.

出版信息

iScience. 2022 Apr 29;25(5):104326. doi: 10.1016/j.isci.2022.104326. eCollection 2022 May 20.

Abstract

Plastic waste imposes a serious problem to the environment and society. Hence, strategies for a circular plastic economy are demanded. One strategy is the engineering of polyester hydrolases toward higher activity for the biotechnological recycling of polyethylene terephthalate (PET). To provide tools for the rapid characterization of PET hydrolases and the detection of degradation products like terephthalic acid (TPA), we coupled a carboxylic acid reductase (CAR) and the luciferase LuxAB. CAR converted TPA into the corresponding aldehydes in , which yielded bioluminescence that not only semiquantitatively reflected amounts of TPA in hydrolysis samples but is suitable as a high-throughput screening assay to assess PET hydrolase activity. Furthermore, the CAR-catalyzed synthesis of terephthalaldehyde was combined with a reductive amination cascade in a one-pot setup yielding the corresponding diamine, suggesting a new strategy for the transformation of TPA as a product obtained from PET biodegradation.

摘要

塑料垃圾给环境和社会带来了严重问题。因此,需要循环塑料经济的策略。其中一种策略是对聚酯水解酶进行工程改造,以提高其活性,用于聚对苯二甲酸乙二酯(PET)的生物技术回收。为了提供快速表征PET水解酶和检测对苯二甲酸(TPA)等降解产物的工具,我们将羧酸还原酶(CAR)与荧光素酶LuxAB偶联。CAR在 中将TPA转化为相应的醛,产生生物发光,这不仅能半定量反映水解样品中TPA的含量,还适合作为评估PET水解酶活性的高通量筛选测定方法。此外,CAR催化对苯二甲醛的合成与一锅法中的还原胺化级联反应相结合,生成相应的二胺,这为将TPA作为PET生物降解产物进行转化提供了一种新策略。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/11a5/9117539/e91e81fc080a/fx1.jpg

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