Mechanisms for the Enhancement of Caproic Acid and H2 Production in Ruminococcaceae Bacterium CPB6 by Fe(II) and Mg(II): Growth and Gene Transcription Analyses.

The production of caproic acid (CA) and hydrogen gas (H2) from organic wastewater is economically attractive. The Ruminococcaceae bacterium CPB6 has demonstrated potential for CA production from lactate-containing wastewater. However, our understanding of the effects of Fe2+ and Mg2+ on the growth and metabolism of strain CPB6 remains limited. Therefore, this study aims to investigate the impact of Fe2+ and Mg2+ on CA and H2 production, as well as on the expression of key genes involved in CA and H2 biosynthesis pathway. The results indicate that Fe2+ positively affects cell proliferation and H2 production while minimally impacting CA production. The highest levels of H2 production were achieved with the addition of 200 mg/L Fe2+. Conversely, Mg2+ significantly enhances CA and H2 production, with the optimal yield observed in a medium enriched with 300 mg/L Mg2+. Reverse transcription quantitative PCR (RT-qPCR) analysis reveals that Fe2+ promotes the expression of the hydrogenase gene, whereas Mg2+ has a negligible effect on hydrogenase expression. Notably, Fe2+ and Mg2+ inhibit the expression of key genes involved in CA synthesis. These findings suggest that Fe2+ enhances H2 production by boosting cell biomass and the expression of the hydrogenase gene, whereas Mg2+ improves CA and H2 production primarily by increasing cell biomass rather than influencing the expression of functional genes involved in CA biosynthesis.