Light-driven methanol utilization in Escherichia coli for malate production

Abstract

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.

Keywords

Methanol utilization / Malate / Light-driven / ATP supply / E. coli

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Funding

the Project of Science and Technology Innovation Team of Hunan Province(2021RC4063)

the Outstanding Youth Project of Hunan Provincial Department of Education(24B0252)

RIGHTS & PERMISSIONS

Jiangnan University

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