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<p>The exploration of sustainable biofuels has emerged as a competitive alternative to petroleum. Isoprenol, a C5 isoprenoid, has gained prominence as a potential substitute for gasoline with a high octane number and combustion efficiency. Consequently, microbial production of isoprenol has been extensively explored. Given that the isoprenol-production pathway is initiated by a stepwise condensation of three molecules of acetyl-CoA, ensuring the acetyl-CoA availability has been crucial for efficient isoprenol production. Prior research indirectly addressed this challenge by cultivating cells in nutrient-rich media, which might enhance the production cost. Alternatively, the adoption of precise acetyl-CoA redirection can be a viable strategy for attaining efficient production. We propose a strategy based on the precise flux redirection of acetyl-CoA for efficient production of isoprenol in minimal media. Firstly, we established the isoprenol-production pathway via amplification of the downstream pathway for mevalonate conversion into isoprenol. Subsequently, we endeavored to simultaneously regulate the expression of <i>mvaE</i> and <i>mvaS</i> to optimize the acetyl-CoA availability. The expression-optimized BI02 strain demonstrated a remarkable increase of 1.90-fold in isoprenol production compared to the BI01 strain with amplified two genes. In addition, the BI02 strain even outperformed the BI01 strain, even when the additional nutrient was added. In a fed-batch fermentation using a bioreactor, the BI02 strain produced 390.4&#xa0;mg/L of isoprenol. Overall, these results suggest that the redirection of acetyl-CoA, particularly the precise expression optimization of <i>mvaE</i> and <i>mvaS</i>, can be an alternative strategy for efficient production instead of utilizing additional supplements.</p>

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Efficient isoprenol production in Escherichia coli via acetyl-CoA redirection

  • 期刊论文

  • 发表日期：

    2025年5月29日

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  • Kumyoung Jeung,
  • Seungjin Kim,
  • Myung Hyun Noh,
  • Gyoo Yeol Jung

摘要

The exploration of sustainable biofuels has emerged as a competitive alternative to petroleum. Isoprenol, a C5 isoprenoid, has gained prominence as a potential substitute for gasoline with a high octane number and combustion efficiency. Consequently, microbial production of isoprenol has been extensively explored. Given that the isoprenol-production pathway is initiated by a stepwise condensation of three molecules of acetyl-CoA, ensuring the acetyl-CoA availability has been crucial for efficient isoprenol production. Prior research indirectly addressed this challenge by cultivating cells in nutrient-rich media, which might enhance the production cost. Alternatively, the adoption of precise acetyl-CoA redirection can be a viable strategy for attaining efficient production. We propose a strategy based on the precise flux redirection of acetyl-CoA for efficient production of isoprenol in minimal media. Firstly, we established the isoprenol-production pathway via amplification of the downstream pathway for mevalonate conversion into isoprenol. Subsequently, we endeavored to simultaneously regulate the expression of mvaE and mvaS to optimize the acetyl-CoA availability. The expression-optimized BI02 strain demonstrated a remarkable increase of 1.90-fold in isoprenol production compared to the BI01 strain with amplified two genes. In addition, the BI02 strain even outperformed the BI01 strain, even when the additional nutrient was added. In a fed-batch fermentation using a bioreactor, the BI02 strain produced 390.4 mg/L of isoprenol. Overall, these results suggest that the redirection of acetyl-CoA, particularly the precise expression optimization of mvaE and mvaS, can be an alternative strategy for efficient production instead of utilizing additional supplements.