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使用聚呋喃酸乙烯酯可溶性支架对聚对苯二甲酸乙二酯脂肪酶进行定向进化的酶标测

Enzyme Benchmarking with Polyethylene Furanoate Soluble Scaffolds for Directed Evolution of PEFases.

作者信息

Dolz Mikel, Monterrey Dianelis T, Quartinello Felice, de Santos Patricia Gomez, Mateljak Ivan, Pellis Alessandro, Guebitz Georg, Viña-González Javier, Alcalde Miguel

机构信息

EvoEnzyme S.L., Parque Científico de Madrid, Cantoblanco, 28049 Madrid, Spain.

Department of Biocatalysis, Institute of Catalysis, CSIC, Marie Curie 2, Cantoblanco, 28049 Madrid, Spain.

出版信息

ACS Omega. 2024 Nov 1;9(45):45633-45640. doi: 10.1021/acsomega.4c09053. eCollection 2024 Nov 12.

DOI:10.1021/acsomega.4c09053
PMID:39554451
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11561765/
Abstract

Plastic waste is a major threat in our industrialized world and is driving research into bioplastics. The success of biobased polyethylene furanoate (PEF) as a viable alternative to polyethylene terephthalate (PET) of fossil origin will depend on designing effective enzymes to break it down, aiding its recycling. Here, a panel of fungal and bacterial cutinases were functionally expressed in a tandem yeast expression system based on and . The activity of the enzyme panel was tested with soluble PEF model scaffolds, observing a correlation with the degradation of real PEF powder. A high-throughput colorimetric screening assay based on the PEF scaffold diethyl furan-2,5-dicarboxylate was developed, establishing the basis for future directed evolution campaigns of PEFases.

摘要

塑料垃圾是我们工业化世界中的一个重大威胁，正推动着对生物塑料的研究。生物基聚呋喃二甲酸乙二醇酯（PEF）作为化石来源的聚对苯二甲酸乙二酯（PET）的可行替代品，其成功将取决于设计有效的酶来分解它，以帮助其回收利用。在此，一组真菌和细菌角质酶在基于[具体内容缺失]和[具体内容缺失]的串联酵母表达系统中进行了功能表达。用可溶性PEF模型支架测试了该酶组的活性，观察到与实际PEF粉末的降解存在相关性。开发了一种基于PEF支架2,5-二羧酸二乙酯的高通量比色筛选测定法，为未来PEF酶的定向进化活动奠定了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c558/11561765/179b580ba935/ao4c09053_0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c558/11561765/0e35dde5f16a/ao4c09053_0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c558/11561765/180218ae9751/ao4c09053_0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c558/11561765/73fca7d5a133/ao4c09053_0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c558/11561765/41269402ef35/ao4c09053_0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c558/11561765/676f9c02115e/ao4c09053_0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c558/11561765/179b580ba935/ao4c09053_0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c558/11561765/0e35dde5f16a/ao4c09053_0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c558/11561765/180218ae9751/ao4c09053_0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c558/11561765/73fca7d5a133/ao4c09053_0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c558/11561765/41269402ef35/ao4c09053_0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c558/11561765/676f9c02115e/ao4c09053_0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c558/11561765/179b580ba935/ao4c09053_0005.jpg

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本文引用的文献

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Efficient Depolymerization of Poly(ethylene 2,5-furanoate) Using Polyester Hydrolases.利用聚酯水解酶高效解聚聚（2,5-呋喃二甲酸乙二酯）
ACS Sustain Chem Eng. 2024 Jun 20;12(26):9658-9668. doi: 10.1021/acssuschemeng.4c00915. eCollection 2024 Jul 1.
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Enzymes' Power for Plastics Degradation.酶在塑料降解方面的作用。
Chem Rev. 2023 May 10;123(9):5612-5701. doi: 10.1021/acs.chemrev.2c00644. Epub 2023 Mar 14.
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A Dual Fluorescence Assay Enables High-Throughput Screening for Poly(ethylene terephthalate) Hydrolases.一种双荧光测定法可实现对聚对苯二甲酸乙二酯水解酶的高通量筛选。
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A flexible kinetic assay efficiently sorts prospective biocatalysts for PET plastic subunit hydrolysis.一种灵活的动力学分析方法能够有效地筛选出用于聚对苯二甲酸乙二酯（PET）塑料亚基水解的潜在生物催化剂。
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