Extracellular electron transfer in fermentative bacterium Anoxybacter fermentans DY22613 isolated from deep-sea hydrothermal sulfides.

Dissimilatory Fe(III)-reducing bacteria (DIRBs) could reduce extracellular Fe(III) to Fe(II) via extracellular electron transfer (EET), playing an important role in biogeochemical cycling of Fe(III). Previous studies have noted the key role of multi-heme c-type cytochromes (MHCs) involved in EET by respiratory-type DIRBs, and proposed indirect electron transfer through the use of redox electron shuttles (e.g., flavins) or Fe(III)-chelation. However, knowledge about the EET of fermentative DIRBs was vitally scarce. Here, Anoxybacter fermentans DY22613 is a typical fermentative DIRB isolated from deep-sea hydrothermal sulfides, and it could utilize soluble Fe(III)-citrate and solid Fe(III)-bearing minerals as extracellular electron acceptors. Unlike respiratory-type DIRBs that utilize MHCs, this strain lacked MHCs to mediate EET. Besides, it did not adopt Fe(III)-chelation to mediate indirect EET. Nonetheless, genes encoding biosynthesis pathway of redox molecules (e.g., flavins) were found in its genome and their gene expression was up-regulated with Fe(III) reduction, suggesting redox molecules may mediate indirect EET by this strain. Subsequent physiological and biochemical tests further demonstrated endogenous riboflavin acted as main electron shuttles to mediate indirect EET by this strain, and menaquinone, indole played an assistant role in this process. Besides, this strain could employ exogenous humic acids to facilitate indirect EET. The mode of exogenous and endogenous redox molecules to co-mediate indirect EET by fermentative A. fermentans DY22613, expands our knowledge about EET of fermentative DIRBs, and would contribute to better understand its ecological role in the biogeochemistry cycle of iron.