Work

PerR Deletion Enhances Oxygen Tolerance and Butanol/Acetone Production in a Solvent-Degenerated Clostridium beijerinckii Strain DS

Year: 2025

Type: article

Abstract: The industrial potential of Clostridium beijerinckii for acetone–butanol–ethanol (ABE) fermentation is limited by oxygen sensitivity and suboptimal solvent productivity. Peroxide repressor (PerR), a key negative regulator protein, is reported to suppress the oxidative stress defense system in anaerobic clostridia, leading to poor survival of bacteria under aerobic conditions. However, the regulatory mechanism underlying this phenomenon remains unclear. This study demonstrates that targeted deletion of perR (Cbei_1336) in the solvent-deficient strain C. beijerinckii DS confers robust oxygen tolerance and enhances ABE fermentation performance. The engineered perR mutant exhibited unprecedented aerobic growth under atmospheric oxygen (21% O2), achieving a (3.79 ± 0.09)-fold increase in biomass accumulation, a (2.84 ± 0.12)-fold improvement in glucose utilization efficiency, a (57.23 ± 0.01)-fold elevation in butanol production, and a (32.78 ± 0.02)-fold amplification in acetone output compared to the parental strain. Transcriptomic analysis revealed that perR knockout simultaneously upregulated oxidative defense systems and activated ABE pathway-related genes. This genetic rewiring redirected carbon flux from acidogenesis to solventogenesis, yielding a (9.64 ± 0.90)-fold increase in total solvent titer (15.61 ± 0.89 vs. 1.62 ± 0.12 g/L) and a (2.71 ± 0.04)-fold rise in volumetric productivity (0.19 ± 0.01 vs. 0.07 ± 0.01 g/L/h). Our findings establish PerR as a master regulator of both oxygen resilience and metabolic reprogramming, providing a scalable engineering strategy for industrial oxygen-tolerant ABE bioprocessing toward low-cost biobutanol production.

Source: Fermentation

DOI: 10.3390/fermentation11090526