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<p>Benzoic acid, an aromatic compound conventionally obtained from coal tar, can also be produced via microbial biotransformation. The shikimate pathway of <i>Escherichia coli</i> provides a route for the biosynthesis of aromatic acids, with its intermediates serving as valuable starting materials for the synthesis of benzoic acid derivatives. Here, we report the <i>E. coli</i>-based synthesis of three benzoic acid derivatives: 3,4-dihydroxybenzoic acid (protocatechuic acid, 3,4-DHBA), gallic acid (GA), and β-glucogallin. The enzyme QuiC was used to catalyze the conversion of 3-dehydroshikimate to 3,4-DHBA. For GA production, a screening of four <i>pobA</i> mutants was conducted to identify the most efficient mutant. A grape-derived uridine diphosphate-dependent glucosyltransferase (UDP-GT) was utilized for the glucosylation of GA to β-glucogallin. To improve the production titers of 3,4-DHBA and GA, a shikimate pathway gene module and a specifically engineered <i>E. coli</i> mutant were employed, resulting in the accumulation of 451.3&#xa0;mg/L 3,4-DHBA and 123.4&#xa0;mg/L GA. β-Glucogallin synthesis was achieved through a stepwise process, wherein one <i>E. coli</i> strain produced GA, and its clarified culture medium was subsequently used by a second <i>E. coli</i> strain for β-glucogallin formation. Optimization of the ratio between the GA-containing supernatant and the second cell culture led to a β-glucogallin yield of 118.5&#xa0;mg/L.</p>

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Biosynthesis of three benzoic acid derivatives in Escherichia coli

  • 期刊论文

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  • 发表日期：

    2025年8月13日

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  • Hyun-Ah Kim,
  • Hyun Ji Kang,
  • Joong-Hoon Ahn

摘要

Benzoic acid, an aromatic compound conventionally obtained from coal tar, can also be produced via microbial biotransformation. The shikimate pathway of Escherichia coli provides a route for the biosynthesis of aromatic acids, with its intermediates serving as valuable starting materials for the synthesis of benzoic acid derivatives. Here, we report the E. coli-based synthesis of three benzoic acid derivatives: 3,4-dihydroxybenzoic acid (protocatechuic acid, 3,4-DHBA), gallic acid (GA), and β-glucogallin. The enzyme QuiC was used to catalyze the conversion of 3-dehydroshikimate to 3,4-DHBA. For GA production, a screening of four pobA mutants was conducted to identify the most efficient mutant. A grape-derived uridine diphosphate-dependent glucosyltransferase (UDP-GT) was utilized for the glucosylation of GA to β-glucogallin. To improve the production titers of 3,4-DHBA and GA, a shikimate pathway gene module and a specifically engineered E. coli mutant were employed, resulting in the accumulation of 451.3 mg/L 3,4-DHBA and 123.4 mg/L GA. β-Glucogallin synthesis was achieved through a stepwise process, wherein one E. coli strain produced GA, and its clarified culture medium was subsequently used by a second E. coli strain for β-glucogallin formation. Optimization of the ratio between the GA-containing supernatant and the second cell culture led to a β-glucogallin yield of 118.5 mg/L.