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<p>(−)-α-bisabolol is a sesquiterpene compound found in various plants, with broad applications in pharmaceuticals, cosmetics, and personal care products due to its diverse pharmacological properties. Recombinant <i>Escherichia coli</i> offers an efficient and sustainable platform for its production. To further advance its industrial-scale synthesis, we enhanced the expression and catalytic activity of the key enzyme bisabolol synthase (CcBOS) and optimized the metabolic pathways of the host chassis. In this study, CcBOS from <i>Cynara cardunculus</i> was expressed in a previously engineered sesquiterpene-producing <i>E. coli</i> strain. By adjusting the fermentation temperature to 25&#xa0;°C, the (−)-α-bisabolol titer reached 1.12&#xa0;g/L in shake flask cultures. Protein engineering strategies—including optimization of the N-terminal sequence, increasing polar amino acid content, modifying non-conserved residues, and mutating histidine residues near the active site—significantly improved the solubility and catalytic efficiency of CcBOS. The triple mutant T258S/I364N/H479A increased (−)-α-bisabolol production by 154.5%. Metabolic engineering efforts, such as deleting <i>atoDA</i> to reduce fatty acid synthesis and attenuating <i>gltA</i> expression to limit tricarboxylic acid (TCA) cycle competition, further enhanced the titer by 63.1%. Integrating protein and metabolic engineering approaches resulted in a final titer of 3.08&#xa0;g/L in shake flasks. When scaled up to a 5&#xa0;L bioreactor, the titer reached 7.41&#xa0;g/L, representing a high production of (−)-α-bisabolol using glucose as the sole carbon source. This study provides a novel integrated approach for the efficient microbial synthesis of (−)-α-bisabolol.</p>

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Evaluation of protein and metabolic engineering strategies on (−)-α-bisabolol production by Escherichia coli

  • 期刊论文

  • 发表日期：

    2026年5月21日

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  • Jiayi Ma,
  • Rong Fang,
  • Xiaoyuan Yue,
  • Hui Jiang,
  • Qixuan Kong,
  • Tiankuo Jiang,
  • Jiale Yu,
  • Chang Zhou,
  • Yuanyuan Xia,
  • Wei Shen,
  • Haili Liu,
  • Yaqin Shen,
  • Haiquan Yang,
  • Li Zhou,
  • Xianzhong Chen

摘要

(−)-α-bisabolol is a sesquiterpene compound found in various plants, with broad applications in pharmaceuticals, cosmetics, and personal care products due to its diverse pharmacological properties. Recombinant Escherichia coli offers an efficient and sustainable platform for its production. To further advance its industrial-scale synthesis, we enhanced the expression and catalytic activity of the key enzyme bisabolol synthase (CcBOS) and optimized the metabolic pathways of the host chassis. In this study, CcBOS from Cynara cardunculus was expressed in a previously engineered sesquiterpene-producing E. coli strain. By adjusting the fermentation temperature to 25 °C, the (−)-α-bisabolol titer reached 1.12 g/L in shake flask cultures. Protein engineering strategies—including optimization of the N-terminal sequence, increasing polar amino acid content, modifying non-conserved residues, and mutating histidine residues near the active site—significantly improved the solubility and catalytic efficiency of CcBOS. The triple mutant T258S/I364N/H479A increased (−)-α-bisabolol production by 154.5%. Metabolic engineering efforts, such as deleting atoDA to reduce fatty acid synthesis and attenuating gltA expression to limit tricarboxylic acid (TCA) cycle competition, further enhanced the titer by 63.1%. Integrating protein and metabolic engineering approaches resulted in a final titer of 3.08 g/L in shake flasks. When scaled up to a 5 L bioreactor, the titer reached 7.41 g/L, representing a high production of (−)-α-bisabolol using glucose as the sole carbon source. This study provides a novel integrated approach for the efficient microbial synthesis of (−)-α-bisabolol.