Presentation Information
[P03-339]Conjugated Polymer-Microbe Interactions Trigger Regulatory Reprogramming for Enhanced Bioelectrocatalysis
○Yuanmei Liang1,3,4,5, Ohayon David2, Zexuan Wang1,3,4,5, Jee Loon Foo1,3,4,5, Glenn Quek6, Samantha R McCuskey7, Zhongxin Chen2, Matthew Wook Chang1,3,4,5, Guillermo C Bazan2,7 (1. NUS Synthetic Biology for Clinical and Technological Innovation (SynCTI) (Singapore), 2. Institute for Functional Intelligent Materials, National University of Singapore (Singapore), 3. Synthetic Biology Translational Research Program, Yong Loo Lin School of Medicine, National University of Singapore (Singapore), 4. Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, (Singapore), 5. National Centre for Engineering Biology (NCEB) (Singapore), 6. Departments of Chemistry and Chemical & Biomolecular Engineering, National University of Singapore (Singapore), 7. Institute for Digital Molecular Analytics and Science (IDMxS), Nanyang Technological University (Singapore))
Keywords:
microbial electrosynthesis,poly(benzodifurandione) (PBFDO),biohybrid interface,extracellular electron transfer,metabolomic reprogramming,Shewanella oneidensis MR-1
[Purpose]
Abiotic redox-active materials are increasingly used to interface with electroactive microorganisms, but whether they actively reshape microbial regulation remains unclear. This study aimed to determine how the electron-transporting n-type conjugated polymer poly(benzodifurandione) (PBFDO) affects inward extracellular electron transfer (EET) and cellular physiology in Shewanella oneidensis MR-1 under cathodic conditions.
[Method]
We coupled S. oneidensis MR-1 with PBFDO-coated carbon electrodes and evaluated electron uptake and fumarate reduction under cathodic operation. Electrochemical measurements were integrated with transcriptomic and metabolomic analyses to examine changes in electron-transfer pathways, central metabolism, biofilm-related responses, and intracellular redox and energy states. Polymer-responsive small RNAs (sRNAs) were also identified, and selected sRNAs were experimentally delivered into control cells to test whether they could reproduce polymer-associated phenotypes.
[Results]
PBFDO-coated electrodes markedly enhanced electron uptake and fumarate reduction, producing over two orders of magnitude higher current density and up to a twelvefold increase in succinate production compared with polymer-free controls. Multi-omics analysis showed that these improvements resulted from coordinated physiological reprogramming rather than passive conductivity alone. PBFDO selectively strengthened the Mtr-CymA electron-transfer conduit, central carbon metabolism, and ATP synthesis, while promoting surface adhesion and biofilm formation and suppressing membrane protein folding pathways. Metabolomic profiling further revealed rerouting of tricarboxylic acid cycle flux toward fumarate reduction, together with elevated NADH/NAD+ and ATP/ADP ratios. Importantly, a distinct class of polymer-responsive sRNAs was identified, indicating a post-transcriptional regulatory layer associated with the synthetic interface. Delivery of individual sRNAs into control cells partially recapitulated the polymer-induced phenotype, increasing current output by up to threefold and altering intracellular redox balance.
[Consideration]
These findings suggest that the polymer-cell interface does more than improve interfacial conductivity; it actively engages bacterial regulatory networks at both transcriptional and post-transcriptional levels. The identification of polymer-responsive sRNAs reveals a previously unrecognized mechanism linking synthetic materials to microbial regulatory control, metabolism, and electron transfer.
[Conclusion]
PBFDO forms a biohybrid interface that enhances inward EET and fumarate reduction in S. oneidensis MR-1 through coordinated metabolic and regulatory reprogramming. The newly revealed interface-sRNA-metabolism axis provides a conceptual framework for microbial electrosynthesis and suggests new design principles for engineering functional biohybrid interfaces.
Abiotic redox-active materials are increasingly used to interface with electroactive microorganisms, but whether they actively reshape microbial regulation remains unclear. This study aimed to determine how the electron-transporting n-type conjugated polymer poly(benzodifurandione) (PBFDO) affects inward extracellular electron transfer (EET) and cellular physiology in Shewanella oneidensis MR-1 under cathodic conditions.
[Method]
We coupled S. oneidensis MR-1 with PBFDO-coated carbon electrodes and evaluated electron uptake and fumarate reduction under cathodic operation. Electrochemical measurements were integrated with transcriptomic and metabolomic analyses to examine changes in electron-transfer pathways, central metabolism, biofilm-related responses, and intracellular redox and energy states. Polymer-responsive small RNAs (sRNAs) were also identified, and selected sRNAs were experimentally delivered into control cells to test whether they could reproduce polymer-associated phenotypes.
[Results]
PBFDO-coated electrodes markedly enhanced electron uptake and fumarate reduction, producing over two orders of magnitude higher current density and up to a twelvefold increase in succinate production compared with polymer-free controls. Multi-omics analysis showed that these improvements resulted from coordinated physiological reprogramming rather than passive conductivity alone. PBFDO selectively strengthened the Mtr-CymA electron-transfer conduit, central carbon metabolism, and ATP synthesis, while promoting surface adhesion and biofilm formation and suppressing membrane protein folding pathways. Metabolomic profiling further revealed rerouting of tricarboxylic acid cycle flux toward fumarate reduction, together with elevated NADH/NAD+ and ATP/ADP ratios. Importantly, a distinct class of polymer-responsive sRNAs was identified, indicating a post-transcriptional regulatory layer associated with the synthetic interface. Delivery of individual sRNAs into control cells partially recapitulated the polymer-induced phenotype, increasing current output by up to threefold and altering intracellular redox balance.
[Consideration]
These findings suggest that the polymer-cell interface does more than improve interfacial conductivity; it actively engages bacterial regulatory networks at both transcriptional and post-transcriptional levels. The identification of polymer-responsive sRNAs reveals a previously unrecognized mechanism linking synthetic materials to microbial regulatory control, metabolism, and electron transfer.
[Conclusion]
PBFDO forms a biohybrid interface that enhances inward EET and fumarate reduction in S. oneidensis MR-1 through coordinated metabolic and regulatory reprogramming. The newly revealed interface-sRNA-metabolism axis provides a conceptual framework for microbial electrosynthesis and suggests new design principles for engineering functional biohybrid interfaces.
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