Engineering energy-robust Clostridium autoethanogenum via integrated nanotechnology and transcription-guided metabolic reprogramming.

Microbial conversion of CO and CO2 offers a promising route to C1 gas valorization, but acetogenic syngas fermentation remains constrained by bioenergetic bottlenecks. To address this, a biohybrid system integrating bovine serum albumin-stabilized gold nanoparticles with Clostridium autoethanogenum was constructed, increasing intracellular ATP levels, modulating redox balance, and promoting cell growth under illumination. Transcriptomic analysis of this system systematically identified key energy complexes and positive regulators associated with enhanced cellular energy metabolism. Based on these elements, a Push-Pull-Block strategy was implemented to generate the target strain CAB181, which increased OD600-normalized production of ethanol, acetate, and total acids and alcohols by 46.5%, 35.1%, and 34.2%, respectively. Multiomics analysis confirmed molecular-level metabolic reprogramming in CAB181. This work provides an effective strategy to improve the bioenergetic robustness and fermentation performance of C. autoethanogenum-based cell factories, and offers a reference framework for future engineering of other acetogens.