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<p>Trans-anethole stands as a significant natural compound, finding extensive application as a flavoring ingredient across the food, cosmetics, perfumery, and pharmaceutical sectors. Trans-anethole also offers a variety of positive health benefits to humans, including anti-cancer, anti-inflammatory, anti-diabetic, immunomodulatory, and neuroprotective effects. In this study, we constructed a <i>de novo</i> trans-anethole pathway in <i>E. coli</i> by exploring the reduction of p-coumaric acid to p-coumaryl alcohol through carboxylate reductase and endogenous aldehyde reductase. First, an engineered <i>Escherichia coli</i> was created that could biosynthesise trans-anethole from glucose by the co-expression of <i>aroG</i><sup>fbr</sup>, <i>tyrA</i><sup>fbr</sup>,<i>TAL</i>, <i>CAR</i>, <i>Sfp</i>, <i>ATF1</i>, <i>LtPPS1</i> and <i>AIMT1</i>. Second, by increasing the availability of SAM, we further increased the production of trans-anethole to 125&#xa0;mg/L. Subsequently, the genes for transketolase I <i>tktA</i> and phosphoenolpyruvate synthase <i>ppsA</i> were further overexpressed to increase the availability of erythrose-4-phosphate and phosphoenolpyruvate, resulting in an increase in trans-anethole production to 223&#xa0;mg/L. Finally, by knocking out the <i>pheA</i> gene to block the competitive pathway, the titer of trans-anethole was increased to 356&#xa0;mg/L.</p>

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Carboxylic acid reductase-dependent biosynthesis of trans-anethole in engineered Escherichia coli

  • 期刊论文

  • 发表日期：

    2026年1月8日

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  • Qiuli Wang,
  • Shaoting Wu,
  • Shuxian Ma,
  • Hong Pan,
  • Daoyi Guo

摘要

Trans-anethole stands as a significant natural compound, finding extensive application as a flavoring ingredient across the food, cosmetics, perfumery, and pharmaceutical sectors. Trans-anethole also offers a variety of positive health benefits to humans, including anti-cancer, anti-inflammatory, anti-diabetic, immunomodulatory, and neuroprotective effects. In this study, we constructed a de novo trans-anethole pathway in E. coli by exploring the reduction of p-coumaric acid to p-coumaryl alcohol through carboxylate reductase and endogenous aldehyde reductase. First, an engineered Escherichia coli was created that could biosynthesise trans-anethole from glucose by the co-expression of aroGfbr, tyrAfbr,TAL, CAR, Sfp, ATF1, LtPPS1 and AIMT1. Second, by increasing the availability of SAM, we further increased the production of trans-anethole to 125 mg/L. Subsequently, the genes for transketolase I tktA and phosphoenolpyruvate synthase ppsA were further overexpressed to increase the availability of erythrose-4-phosphate and phosphoenolpyruvate, resulting in an increase in trans-anethole production to 223 mg/L. Finally, by knocking out the pheA gene to block the competitive pathway, the titer of trans-anethole was increased to 356 mg/L.