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<p>As a representative one-carbon compound, methanol has emerged as an ideal alternative substrate for biomanufacturing applications, owing to its abundant availability and low production cost. However, the challenge faced by heterotrophic microbial cells in utilizing methanol to synthesize chemicals is the insufficient energy driving force. Here, a light-driven ATP supply system was constructed to enhance the efficiency of malate production using methanol as a substrate in <i>E. coli</i>. Firstly, a methanol-utilizing malate biosynthesis pathway was constructed in <i>E. coli</i>, resulting in strain ZY-1 with malate production and methanol consumption reaching 9.79&#xa0;g/L and 15.21 mM, respectively. Then, a light-driven ATP supply system was constructed through intracellular synthesis of MgP molecules, and the ATP supply efficiency was further optimized by co-expression of genes <i>nadD</i> and <i>ubiV</i>, resulting in a 49.80% increase in the intracellular ATP content of strain ZY-5. Finally, light-driven methanol utilization for malate production was achieved by developing iLCRC strategy, resulting in malate production and methanol consumption of strain ZY-5 reaching 17.18&#xa0;g/L and 68.55 mM, respectively. This light-driven ATP supply system offers novel insights into improving the future resource utilization of one-carbon compounds.</p>

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Light-driven methanol utilization in Escherichia coli for malate production

  • 期刊论文

  • 发表日期：

    2025年8月8日

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  • Zongyu Yang,
  • Xiaoxuan Guo,
  • Pei Zhou,
  • Xujie Dong,
  • Xiaoling Wang,
  • Gao-Qiang Liu,
  • Tian Tong

摘要

As a representative one-carbon compound, methanol has emerged as an ideal alternative substrate for biomanufacturing applications, owing to its abundant availability and low production cost. However, the challenge faced by heterotrophic microbial cells in utilizing methanol to synthesize chemicals is the insufficient energy driving force. Here, a light-driven ATP supply system was constructed to enhance the efficiency of malate production using methanol as a substrate in E. coli. Firstly, a methanol-utilizing malate biosynthesis pathway was constructed in E. coli, resulting in strain ZY-1 with malate production and methanol consumption reaching 9.79 g/L and 15.21 mM, respectively. Then, a light-driven ATP supply system was constructed through intracellular synthesis of MgP molecules, and the ATP supply efficiency was further optimized by co-expression of genes nadD and ubiV, resulting in a 49.80% increase in the intracellular ATP content of strain ZY-5. Finally, light-driven methanol utilization for malate production was achieved by developing iLCRC strategy, resulting in malate production and methanol consumption of strain ZY-5 reaching 17.18 g/L and 68.55 mM, respectively. This light-driven ATP supply system offers novel insights into improving the future resource utilization of one-carbon compounds.