Background: Microbial electrosynthesis (MES) has been considered a promising platform for carboxylic acids production using homoacetogenic and chain-elongating bacteria. However, this biotechnology is still in its infancy, and many limitations are needed to be transcended, such as low electron transfer efficiency between cathode and microbes.Results: In this study, nano zero-valent iron (NZVI) was employed to improve carboxylic acid production in the MES system. The final accumulated acetate, butyrate, and caproate concentration enhanced from 700.1, 53.6, and 11.8 mg/L to 1.6, 1.1, and 0.7 g/L, with the maximum yield on bicarbonate of 20.7%, 18.7%, and 13.7%, respectively. Besides performing as an additional electron donor, NZVI was also proven to facilitate the formation of electroactive biofilms according to the results of DNA concentration. In addition, it was found that the anaerobic corrosion of NZVI promoted the domination of hydrogen-producing bacteria Paraclostridium, with the increasing relative abundance from 16.78% to 44.18% in the biofilm under open-circuit mode. While under the closed-circuit, the presence of NZVI resulted in significantly higher relative abundances of homoacetogenic bacteria (Terrisporobacter: 20.91%) and chain-elongating bacteria (Clostridium-sensu-stricto-12: 11.8%) in biofilm than in the catholyte (2.18% and 1.63% correspondingly).Conclusions: This study provided the evidence and revealed how NZVI assisted carboxylic acid production in MES and contributes to the application prospects of MES for carbon utilization and energy sustainability.