Patent · US Active

Recombinant Escherichia coli strain for producing succinic acid and construction method thereof

US11976267B2 · kind B2 · utility

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10Claims
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Key dates

Filing dateNov 12, 2020
Grant dateMay 7, 2024
Priority date
Expiry dateOct 7, 2042

Classification

  • Technology area (CPC C)Chemistry; Metallurgy
  • CPC primaryC12R2001/19
  • WIPO fieldBiotechnology
  • WIPO sectorChemistry

Abstract

The invention provides a recombinant Escherichia coli strain for producing succinic acid and a construction method thereof. The by-product encoding genes in the E. coli strain FMME-N-2 are knocked out to obtain the E. coli strain FMME-N-5 (ΔfocA-pflB-ΔldhA-Δpta-ackA); and the phosphoenolpyruvate carboxykinase pck derived from Actinobacillus succinogenes and the phosphite dehydrogenase ptxD derived from Pseudomonas stutzeri were overexpressed. The constructed plasmid pTrcHisA-pck-ptxD was introduced into the expression host E. coli FMME-N-5 (ΔfocA-pflB-ΔldhA-Δpta-ackA), and the cells were screened in a plate containing ampicillin, to obtain an engineered strain E. coli FMME-N-5 (ΔfocA-pflB-ΔldhA-Δpta-ackA)-pck-ptxD that can efficiently produce succinic acid. After fermentation by a two-stage fermentation strategy, the production of succinic acid reaches 137 g/L, the yield of succinic acid is up to 1 g/g glucose, and the space time yield is 1.43 g/L/h, while no by-products of lactic acid and formic acid are accumulated, and the acetic acid content is 1-2 g/L.

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