Biotransformation of ethylene glycol by engineered Escherichia coli

IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Synthetic and Systems Biotechnology Pub Date : 2024-04-11 DOI:10.1016/j.synbio.2024.04.006
Wenlong Yan , Xinhua Qi , Zhibei Cao , Mingdong Yao , Mingzhu Ding , Yingjin Yuan
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Abstract

There has been extensive research on the biological recycling of PET waste to address the issue of plastic waste pollution, with ethylene glycol (EG) being one of the main components recovered from this process. Therefore, finding ways to convert PET monomer EG into high-value products is crucial for effective PET waste recycling. In this study, we successfully engineered Escherichia coli to utilize EG and produce glycolic acid (GA), expecting to facilitate the biological recycling of PET waste. The engineered E. coli, able to utilize 10 g/L EG to produce 1.38 g/L GA within 96 h, was initially constructed. Subsequently, strategies based on overexpression of key enzymes and knock-out of the competing pathways are employed to enhance EG utilization along with GA biosynthesis. An engineered E. coli, characterized by the highest GA production titer and substrate conversion rate, was obtained. The GA titer increased to 5.1 g/L with a yield of 0.75 g/g EG, which is the highest level in the shake flake experiments. Transcriptional level analysis and metabolomic analysis were then conducted, revealing that overexpression of key enzymes and knock-out of the competing pathways improved the metabolic flow in the EG utilization. The improved metabolic flow also leads to accelerated synthesis and metabolism of amino acids.

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工程大肠杆菌对乙二醇的生物转化
为解决塑料废弃物污染问题,人们对 PET 废弃物的生物循环利用进行了广泛研究,乙二醇(EG)是这一过程中回收的主要成分之一。因此,找到将 PET 单体乙二醇转化为高价值产品的方法对于有效回收 PET 废弃物至关重要。在这项研究中,我们成功改造了大肠杆菌,使其能够利用 EG 生产乙醇酸(GA),从而有望促进 PET 废弃物的生物回收利用。最初构建的工程大肠杆菌能在 96 小时内利用 10 克/升的 EG 产生 1.38 克/升的 GA。随后,采用过表达关键酶和敲除竞争途径的策略来提高 EG 的利用率和 GA 的生物合成。经过改造的大肠杆菌具有最高的 GA 产量滴度和底物转化率。GA 滴度增加到 5.1 克/升,EG 产量为 0.75 克/克,这是摇片实验中的最高水平。随后进行了转录水平分析和代谢组学分析,结果表明,关键酶的过度表达和竞争途径的敲除改善了 EG 利用过程中的代谢流。代谢流的改善还导致氨基酸的合成和代谢加快。
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来源期刊
Synthetic and Systems Biotechnology
Synthetic and Systems Biotechnology BIOTECHNOLOGY & APPLIED MICROBIOLOGY-
CiteScore
6.90
自引率
12.50%
发文量
90
审稿时长
67 days
期刊介绍: Synthetic and Systems Biotechnology aims to promote the communication of original research in synthetic and systems biology, with strong emphasis on applications towards biotechnology. This journal is a quarterly peer-reviewed journal led by Editor-in-Chief Lixin Zhang. The journal publishes high-quality research; focusing on integrative approaches to enable the understanding and design of biological systems, and research to develop the application of systems and synthetic biology to natural systems. This journal will publish Articles, Short notes, Methods, Mini Reviews, Commentary and Conference reviews.
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