Use of electricity to direct microbial metabolite production

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Use of electricity to direct microbial metabolite production John M. Pisciotta West Chester University Department of Biology 4th International Conference and Exhibition on Metabolomics and Systems Biology Philadelphia, PA, April 28, 2015 - Talk Objective Discuss recent discoveries detailing how electrical energy, alone or in combination with other factors,

can be used to direct microbial metabolite formation. Touch on the potentials, possibilities and challenges for future research. Highlight the key role Metabolomics can play. - Introduction Organisms have evolved to respond to external stimuli ranging from radiation to chemicals to magnetic fields. Responses can often be measured metabolically. Exposure to stresses like radiation or carcinogens may result in characteristic perturbation of the normal metabolite profile. In humans, induced metabolic changes can help us diagnosis illnesses in microbes they can yield useful products.

Why use Metabolomics? In Biotechnology & Industrial Microbiology the product is often a Metabolite. Pigments (Carotenoids) Nutrients (Omega 3 fatty acids) Antioxidants (Astaxanthin)

Biofuels (Ethanol / Methane) Biosurfactants (Lipopeptides) Antibiotics (Penicillin) Diverse Physicochemical parameters have successfully be used to time, tune & optimize metabolite output. Chemicals pH Light Temperature But what about Electricity as an inducer, and / or energy source?

Bio-Electrochemical System (BES) System that use microbes (and/or cell products) to convert chemical energy to electrical energy, or vice versa, and provide a useful service. BESs use electrode enzymes or cells (usually bacteria) as biocatalysts to drive oxidation & reduction reactions at 2 opposing electrodes. Bioanode and / or Biocathode Metabolomic Methods Used HPLC, GC, MS and potentiostatic techniques are used to study products of mixed or pure

cultures or syntrophic associations. Microbial Fuel Cell 4 e- O2 Substrate CO2 H+ H+ H+ H+ PEM Anode (e- enter circuit)

Anaerobic Air Cathode (e- exit circuit) Aerobic Microbial Fuel Cell 4 e- Metal reducing bacteria O2 Substrate

CO2 H+ H+ H+ H+ PEM Geobacter TEM Cologgi, 2011 Anode (e- enter circuit) Anaerobic Air Cathode (e- exit circuit) Aerobic

Microbial Fuel Cell CO2 H2O PEM H2O Malvankar,et al. 2011 Anode (e- enter circuit) Anaerobic

Air Cathode (e- exit circuit) Aerobic : O2 is e- acceptor > H2O Microbial Electrolysis Cell = H2 Cheng & Logan, 2007, PNAS MECs convert organic wastes (ex. acetate) to usable hydrogen, producing 140%+ more usable energy than electrical energy consumed. Requires applied Voltage CO2 H2 H2 PEM

Anode (e- enter circuit) Anaerobic Cathode (e- exit circuit) Anaerobic : H+ is e- acceptor Electro-methanogenesis Direct Biological Conversion of Electrical Current into Methane by Electro-methanogenesis Cheng et al. 2009. Environ. Sci. Technol. Conversion efficiencies of up to 96% at 1 Volt. Major Advance: Use of BIOCATHODE

(archaea). Implication: Electrically-guided Microbial CO2 fixation Microbial Electrosynthesis Microbial Electrosynthesis: Feeding Microbes Electricity To Convert Carbon Dioxide and Water to Multicarbon Extracellular Organic Compounds Nevin et al., 2010. mBio. Acetogenic bacteria can use electricity to fix CO 2 into organic molecules. Efficiencies of around 80% using Wood Ljungdahl pathway.

Electrosynthesis: The DOE ARPA-E Electrofuels initiative 2010-2013 Enrichment of microbial electrolysis cell (MEC) biocathodes from sediment microbial fuel cell (sMFC) bioanodes. Pisciotta et al., 2012. AEM AEROBIC A) MFC anodes in anaerobic sediment Cathode establish electrogenic biofilm.

Reference B) Electrode inverted to form functional MEC biocathode. Wire Potentiostat A Brush anode Sediment ANAEROBIC

B 1) LSV 2) CV 3) Current Uptake Task 2.3 Milestone Electrotroph Cultivation on CO2 Identification of Electrotrophs

Objectives 1) Demonstrated Transferability 2) Build 16s Clone Libraries Pisciotta etBay al., 2012. AEM Harbor 16s DNA Clone Library: Chesapeake Bay 19 24

Mesorhizobium Rhodococcus Azospirillum Gemmata obscuriglobus Sphingopyxis alaskensis Labrenzia aggregata Endosymbiont of Tevnia Marine actinobacterium Remainder 3 3 3 3

22 10 10 (n = 97) Pure Culture Testing Major Question for Metabolomics The Mechanistic Basis of Electron Uptake? Tremblay & Zhang. 2015.

Frontiers in Microbiology - Summary Electricity can be used for autotrophic conversion of CO2 to useful metabolites. Selective enrichment of electrotrophs from diverse environments is possible. Coupling current with other stimuli (ex. light) may accelerate, expand types of metabolites formed. Metabolomics can help screen for novel electrotroph products, explore voltage effects and determine the electron exchange pathways involved. Acknowledgments JHU Sullivan Lab: Metabolomic GC-MS/MS training.

UMD Baskakov Lab: Metabolomic analysis of pMFC using HPLC w/ PDA detector. PSU Logan Lab: Electrochemical Analysis West Chester University: Hybrid Designs (MEC-PBR) Many thanks to the Meeting Organizers. Let Us Meet Again We welcome you all to our future conferences of OMICS International Please Visit: www.metabolomicsconference.com www.conferenceseries.com http://www.conferenceseries.com/clinical-res

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