Metabolic engineering of Escherichia coli for high-level production of the biodegradable polyester monomer 2-pyrone-4,6-dicarboxylic acid

IF 6.8 1区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Fengli Wu , Shucai Wang , Dan Zhou , Shukai Gao , Guotian Song , Yanxia Liang , Qinhong Wang
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引用次数: 0

Abstract

2-Pyrone-4,6-dicarboxylic acid (PDC), a chemically stable pseudo-aromatic dicarboxylic acid, is a promising building block compound for manufacturing biodegradable polyesters. This study aimed to construct high-performance cell factories enabling the efficient production of PDC from glucose. Firstly, the effective enzymes of the PDC biosynthetic pathway were overexpressed on the chromosome of the 3-dehydroshikimate overproducing strain. Consequently, the one-step biosynthesis of PDC from glucose was achieved. Further, the PDC production was enhanced by multi-copy integration of the key gene PsligC encoding 4-carboxy-2-hydroxymuconate-6-semialdehyde dehydrogenase and co-expression of Vitreoscilla hemoglobin. Subsequently, the PDC production was substantially improved by redistributing the metabolic flux for cell growth and PDC biosynthesis based on dynamically downregulating the expression of pyruvate kinase. The resultant strain PDC50 produced 129.37 g/L PDC from glucose within 78 h under fed-batch fermentation conditions, with a yield of 0.528 mol/mol and an average productivity of 1.65 g/L/h. The findings of this study lay the foundation for the potential industrial production of PDC.

利用大肠杆菌的代谢工程高水平生产可生物降解的聚酯单体 2-吡喃酮-4,6-二羧酸。
2-吡喃酮-4,6-二羧酸(PDC)是一种化学性质稳定的假芳香族二羧酸,是制造生物可降解聚酯的一种前景广阔的基础化合物。本研究旨在构建高性能细胞工厂,以葡萄糖为原料高效生产 PDC。首先,在 3-脱氢莽草酸过量生产菌株的染色体上过表达 PDC 生物合成途径的有效酶。因此,实现了由葡萄糖一步生物合成 PDC。此外,通过多拷贝整合编码 4-羧基-2-羟基琥珀酸-6-半醛脱氢酶的关键基因 PsligC 和共同表达 Vitreoscilla 血红蛋白,提高了 PDC 的产量。随后,在动态下调丙酮酸激酶表达的基础上,通过重新分配细胞生长和 PDC 生物合成的代谢通量,大大提高了 PDC 的产量。在饲料批量发酵条件下,PDC50 菌株在 78 小时内从葡萄糖中产生了 129.37 克/升的 PDC,产量为 0.528 摩尔/摩尔,平均生产率为 1.65 克/升/小时。该研究结果为潜在的 PDC 工业化生产奠定了基础。
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来源期刊
Metabolic engineering
Metabolic engineering 工程技术-生物工程与应用微生物
CiteScore
15.60
自引率
6.00%
发文量
140
审稿时长
44 days
期刊介绍: Metabolic Engineering (MBE) is a journal that focuses on publishing original research papers on the directed modulation of metabolic pathways for metabolite overproduction or the enhancement of cellular properties. It welcomes papers that describe the engineering of native pathways and the synthesis of heterologous pathways to convert microorganisms into microbial cell factories. The journal covers experimental, computational, and modeling approaches for understanding metabolic pathways and manipulating them through genetic, media, or environmental means. Effective exploration of metabolic pathways necessitates the use of molecular biology and biochemistry methods, as well as engineering techniques for modeling and data analysis. MBE serves as a platform for interdisciplinary research in fields such as biochemistry, molecular biology, applied microbiology, cellular physiology, cellular nutrition in health and disease, and biochemical engineering. The journal publishes various types of papers, including original research papers and review papers. It is indexed and abstracted in databases such as Scopus, Embase, EMBiology, Current Contents - Life Sciences and Clinical Medicine, Science Citation Index, PubMed/Medline, CAS and Biotechnology Citation Index.
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