Background <p>Succinic acid (SA) biosynthesis presents a promising sustainable alternative to petrochemical routes, yet industrial potential is often hampered by inefficient CO<sub>2</sub> fixation and redox imbalances.</p> Results <p>To systematically address these bottlenecks, a synergistic metabolic engineering strategy was developed in <i>Actinobacillus succinogenes</i> by co-expressing the endogenous gene of formate dehydrogenase (FdoG) and the heterologous gene of phosphoenolpyruvate carboxylase (PPC). This design established a self-reinforcing metabolic loop where FdoG-mediated formate oxidation generated in situ CO<sub>2</sub> and NADH, which were immediately channeled into the PPC-driven carboxylation pathway. Physiological characterizations, utilizing real-time off-gas analysis, demonstrated that this pathway effectively functions as an endogenous CO<sub>2</sub> generator, decoupling production from external mass transfer limitations. Furthermore, carbon flux distribution analysis confirmed the synergistic efficacy, revealing a 12.11% increase in succinate-directed flux and a significant reduction in byproduct accumulation compared to the wild type. To fully exploit this formate-utilization potential, an optimized fed-batch fermentation with pulsed formate feeding was implemented. Consequently, the engineered strain AS-PF achieved a remarkable succinate titer of 96.64&#xa0;g/L, with a productivity of 1.38&#xa0;g/ (L·h), corresponding to an overall product yield of 0.78&#xa0;g/g based on the total consumed mixed carbon source (glucose and formate).</p> Conclusion <p>This study establishes a sustainable and efficient platform for SA bioproduction by synergistically integrating formate metabolism-driven CO<sub>2</sub> recycling with enhanced biosynthetic pathways, thereby advancing the valorization of C1 compounds.</p>

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Formate-driven in situ CO2 recycling for efficient succinic acid bioproduction in engineered Actinobacillus succinogenes

  • Mingyang Zhao,
  • Li Wang,
  • Yuanming Ye,
  • Yaqin Sun,
  • Zhilong Xiu

摘要

Background

Succinic acid (SA) biosynthesis presents a promising sustainable alternative to petrochemical routes, yet industrial potential is often hampered by inefficient CO2 fixation and redox imbalances.

Results

To systematically address these bottlenecks, a synergistic metabolic engineering strategy was developed in Actinobacillus succinogenes by co-expressing the endogenous gene of formate dehydrogenase (FdoG) and the heterologous gene of phosphoenolpyruvate carboxylase (PPC). This design established a self-reinforcing metabolic loop where FdoG-mediated formate oxidation generated in situ CO2 and NADH, which were immediately channeled into the PPC-driven carboxylation pathway. Physiological characterizations, utilizing real-time off-gas analysis, demonstrated that this pathway effectively functions as an endogenous CO2 generator, decoupling production from external mass transfer limitations. Furthermore, carbon flux distribution analysis confirmed the synergistic efficacy, revealing a 12.11% increase in succinate-directed flux and a significant reduction in byproduct accumulation compared to the wild type. To fully exploit this formate-utilization potential, an optimized fed-batch fermentation with pulsed formate feeding was implemented. Consequently, the engineered strain AS-PF achieved a remarkable succinate titer of 96.64 g/L, with a productivity of 1.38 g/ (L·h), corresponding to an overall product yield of 0.78 g/g based on the total consumed mixed carbon source (glucose and formate).

Conclusion

This study establishes a sustainable and efficient platform for SA bioproduction by synergistically integrating formate metabolism-driven CO2 recycling with enhanced biosynthetic pathways, thereby advancing the valorization of C1 compounds.