Electron shuttle-dependent biofilm formation and biocurrent generation: Concentration effects and mechanistic insights.

Introduction: Electron shuttles (ESs) play a key role in extracellular electron transfer (EET) in Shewanella oneidensis MR-1. However, the quantification relationship between ES concentration, biofilm formation, and biocurrent generation has not been clarified. Methods: In this study, 9,10-anthraquinone-2-sulfonic acid (AQS)-mediated EET and biofilm formation were evaluated at different AQS concentrations in bioelectrochemical systems (BESs) with S. oneidensis MR-1. Results and discussion: Both the biofilm biomass (9- to 17-fold) and biocurrent (21- to 80-fold) were substantially enhanced by exogenous AQS, suggesting the dual ability of AQS to promote both biofilm formation and electron shuttling. Nevertheless, biofilms barely grew without the addition of exogenous AQS, revealing that biofilm formation by S. oneidensis MR-1 is highly dependent on electron shuttling. The biofilm growth was delayed in a BES of 2,000 µM AQS, which is probably because the redundant AQS in the bulk solution acted as a soluble electron acceptor and delayed biofilm formation. In addition, the maximum biocurrent density in BESs with different concentrations of AQS was fitted to the Michaelis-Menten equation (R 2 = 0.97), demonstrating that microbial-catalyzed ES bio-reduction is the key limiting factor of the maximum biocurrent density in BESs. This study provided a fundamental understanding of ES-mediated EET, which could be beneficial for the enrichment of electroactive biofilms, the rapid start-up of microbial fuel cells (MFCs), and the design of BESs for wastewater treatment.

Distinct biofilm formation regulated by different culture media: Implications to electricity generation.

Biofilm of Shewanella oneidensis MR-1 is extensively studied as it can transform organic compounds directly into electricity. Although revealing the biofilm regulation mechanism is crucial for enhancing bio-current, studies regarding the mechanism by which the culture condition affects biofilm formation are still lacking. The biofilm formation of S. oneidensis MR-1 in two typical media with same electron donor was investigated in this study. Bio-electricity increased 1.8 times in medium with phosphate-buffered saline (PBS) than in piperazine-1,4-bisethanesulfonic acid (PIPES). Biofilm total protein has 1.5-fold of difference between two media at day 3, and biofilm structures also differed; a fluffy biofilm with curled cells was formed in medium with PBS, whereas a compact, ordered, and closely attached biofilm was formed in medium with PIPES. Transcriptome studies clarified that the expression of genes beneficial for cell aggregation [e.g., aggA (2.3 fold), bpfA (2.8 fold) and csgB (3.9 fold)] in medium with PIPES was significantly upregulated, thus provided an explanation for the specific biofilm structure. Buffer concentration was proved to be a critical factor impacted cell morphology and current generation. The maximum current density in 30 mM of PBS and PIPES is 165 and 159 µA·cm-2 respectively, but it increased to 327 and 274 µA·cm-2 in 200 mM of PBS and PIPES. This study provides new insights into the mechanism of medium-dependent biofilm regulation, which will be beneficial for developing simple and efficient strategies to enhance bio-electricity generation.