Multilevel Mechanisms of Magnetic Nanoparticles in Enhancing Dark Fermentative Hydrogen Production: From Pure to Mixed Cultures

Dark fermentative hydrogen production is constrained by challenges including low hydrogen yield and operational instability. Magnetic nanoparticles (MNPs) have emerged as promising additives for enhancing biohydrogen production due to their unique physicochemical characteristics, such as high specific surface area, excellent electrical conductivity, and inherent magnetic recyclability. This review systematically compares the enhancement mechanisms of MNPs in two distinct microbial systems: pure cultures and mixed cultures. In pure cultures, MNPs function primarily at the cellular and molecular levels through the following: (1) serving as sustained-release sources of essential metallic cofactors like Fe and Ni to promote hydrogenase synthesis and activation; (2) acting as efficient electron carriers that facilitate intracellular and extracellular electron transfer; and (3) redirecting central carbon metabolism toward high-hydrogen-yield acetate-type fermentation. In mixed cultures, which are more representative of practical applications, MNPs operate at the ecological level through the following: (1) modifying microenvironmental niches to exert selective pressure that enriches hydrogen-producing bacteria, such as Clostridium; (2) forming conductive networks that promote direct interspecies electron transfer and strengthen syntrophic metabolism; and (3) enhancing system robustness via toxin adsorption and pH buffering. Despite promising phenomenological improvements, critical knowledge gaps remain, including unclear structure–activity relationships of MNPs, insufficient quantification of electron transfer pathways, unknown genetic regulatory mechanisms, and overlooked magnetobiological effects. Future research should integrate electrochemical monitoring, multi-omics analyses, and advanced characterization techniques to deepen the mechanistic understanding of nanomaterial–microbe interactions. This review aims to provide theoretical insights and practical strategies for developing efficient and sustainable MNP–microorganism hybrid systems for scalable biohydrogen production.