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Geobiology

MICROBIAL FUEL CELLS (MFCs) Biofuels for energy production and Waste disposal Provost’s Energy Retreat FEEI February 24 & 25, 2006 Ken Nealson Wrigley Professor of Geobiology USC

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PS Bacteria Cyanobacteria Algae & Plants

Geobiology

Energy Flow on Earth
Light Energy (178,000 TW) Animals Fungi Bacteria Biomass Organic C CO2 Geothermal Energy (30 TW) Lithotrophs Reduced Inorganics (organic C) CO2

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Fermentation Metabolism

Geobiology

Biomass

CH3OH CH3CH2OH

3/1 2.5/1

Biofuels – methanol ethanol
Waste (CH4)

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Geobiology
Don’t always have to “win” breaking even might be enough!

Biomass Biofuel Cells
Electricity CO2 Pollution removal water purification industrial water industrial waste

Waste

Biofuel Cell Interruption of the process! (Imagine many other fuels being used by these Microbial Fuel Cells)

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Geobiology

What is a microbial fuel cell? Advantages of MFCs Disadvantages of MFCs State of the Art Challenges Prospects
Demonstrate these with examples of our work when appropriate.

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Geobiology

What is a Microbial Fuel Cell?
Fuel cell with microbe as a catalyst Known since early 1900’s First report of a microbial fuel cell in 1911 (Potter) Take advantage of way life works: Take up fuel, extract electrons electron flow to an acceptor is used to charge a “biological capacitor” charged capacitor used to make biological energy Fuel cell just short circuits this process MFCs come in two types: mediated and mediator-less

Two Types of Microbial Fuel Cells
Mediated Fuel Cell
Load

Mediator-less Fuel Cell
Load

Oxidized Fuel

Oxidant

Oxidized Fuel

Oxidant

Cathode

Cathode

Anode

Anode

Microbe

Microbe Fuel

Reduced Oxidant = Oxidized Mediator = Reduced Mediator = Ion Exchange Membrane

Fuel

Reduced Oxidant = Oxidized carrier molecule = Reduced carrier molecule = Ion Exchange Membrane = Outer membrane electron carriers (i.e. nanowires)

• Almost any microbe can produce electricity with an electron shuttle (innefficient !) • Mediators are mostly phenolic compounds, which are expensive & sometimes toxic • A mediator-less microbial fuel cell is possible if the microbes can give electrons directly to the electrode

• No additional electron shuttle is needed • Few known microbes have this ability • Shewanella & iron reducers do !

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Geobiology

Mediator-less fuel cells take advantage of special bacteria Isolated ~ 15 years ago -- Shewanella, Geobacter, others Iron/manganese reducing bacteria Famous for reducing solid substrates (Fe & Mn oxides) Subsequently found to have enzymes on outside of the cell Unusual for bacteria, but necessary for this reaction Example shown in next slide

Enrichment Culture

Five Days Incubation

Solid Substrate
Pure Culture on MnO2 Breathing Mn oxide!

Biofilm

Microbial Fuel Cell
V
N2 inlet N2 outlet 10 ohm
Pt leads

Air inlet Air outlet

Anode
Injection port for fuel

Cathode

Scale in inches

Pt coated graphite Graphite felt felt electrode electrode Clamp holding ion exchange membrane

Used to evaluate strains in the laboratory Anode – graphite with bacterial catalyst Glass – autoclavable, re-usable Extra ports for electrochemical measurements Cathode platinized graphite (Surya Prakash’s help!)

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Geobiology

Potential advantages of MFCs 1. Catalysts are inexpensive – essentially “free” 2. Catalysts are diverse and robust extreme conditions of pH, Eh, T, salinity, etc. 3.Catalysts are versatile – single type can use wide variety of substrates 4.Catalysts can self repair (proteins, DNA, membranes, etc.)

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Geobiology

More than 50 different Shewanella species known So far, all produce current From ~ 4 oC to 55 oC; wide salinity range 65 different carbon sources Very tough and robust organisms Just the tip of the iceberg of biological diversity (other Fe-reducers are known that grow to 110oC!)

Response to different fuels (Shewanella) Formate
• MR-1 can grow by converting lactate to acetate:
lactate → pyruvate → acetate → CO2
Current (mA) 0.35 0.3 0.25 0.2 0.15 0.1 0.05 0 0 5 10 15 20 25 Lactate Lactate

MFC OD of 0.8

• MR-1 can also use these products to maintain and produce current •Can also switch from one to another with ease:

(1mM)

(1mM)

Form ate

(1mM)

Acetate
0.05 0.045 0.04 0.035 0.03 0.025 0.02 0.015 0.01 0.005 0 0

MFC OD of 0.3
Current (mA)

Lactate Lactate

Acetate

(1mM)

(1mM)

(1mM)
10 20 Tim e (hours) 30 40

0.05 0.045 0.04 0.035 0.03 0.025 0.02 0.015 0.01 0.005 0 0

Succinate

Tim e (Hours)

MFC OD of 0.3

Current (mA)

Succinate Lactate Lactate

(1mM)

(1mM)

(1mM)
10 20 Tim e (hours) 30 40

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Geobiology

Potential disadvantages of MFCs
1. Current density is low 2. Difficult to run and maintain 3. Sensitive to breakdown and decay

Almost certainly all these “disadvantages” are built on misconceptions These arise from use of mediated MFCs

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State of the art:

Geobiology

1. Many bacteria now known that produce current in mediator-less MFCs 2. Mechanism of current production not understood 3. Current densities are getting into the range of interest – mA/cm2 (wide range of abilities) 4. Interesting development has to do with microbial consortia – current density is always higher

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Geobiology

Chang et al., 2006, Electrochemically active bacteria (EAB) and mediator-less microbial fuel cells. J. Microbiol. Biotechnol. 16:163-177. Power densities range from: 16 to 4,300 mW/m2 I have a PDF of this I will send to anyone who wants – reviews much of what I have said today.

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Geobiology

Challenges to be addressed: 1. How do they work? Mechanisms? genetic and genomic approaches 2. Physiology of the cells – interface with FC biofilms, etc. 3. Microbial communities and consortia enrichment cultures

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Geobiology

9 mutants that knock out ability to produce current 4 are involved with iron reduction 5 are not – 3 mutants that increase current production all of these are cytochromes leading to other termini Several regulatory mutants that increase or decrease the level of current production

Table 1. Electrochemical activities of Shewanella oneidensis MR-1 and its mutants
Strain No. 1 2 3 4 5 6 7 8 9 10 11 Gene Growth1 on Lactate/Fumarate Max. current (μA) Coulomb3 (C) CV4 test

ΔluxSrif ΔmtrA ΔomcA ΔhydB ΔhydB and ΔhydA ΔhydA ΔtatC Δmpw Δfur Δcrp
Wild type

+++ +++ +++ +++ +++ +++ + +++ ++ + +++ +++2

65.0±6.1 7.3±0.5 4.6±0.2 61.0±15.4 66.3±17.9 54.0±10.0 15.3±2.9 48.0±7.2 26.0±2.0 19.0±6.6 68.0±7.8 5.0±0.1

2.53±0.25 0.42±0.09 0.32±0.01 2.63±0.37 2.94±0.48 2.53±0.72 0.97±0.16 2.15±0.09 1.30±0.25 1.11±0.36 2.56±0.18 -

++ + +/+ + + ++ + + + + -

12

E.coli

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Geobiology

How do the catalysts work? Mutant screening genome of Shewanella has been sequenced use this information to make directed mutants mutant analysis identifies those genes coding for proteins involved with current prod. so far great success using this approach

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Geobiology

Understanding the catalyst: Role of attachment, biofilms, connections No doubt of catalytic ability Question of how to control and direct it This is issue of physiology of cells:

Shewanella oneidensis MR-1 biofilm current production
1.2

1.2

MR-1 Biofilm on Anode (4 day growth)
1

Injection of planktonic cells (OD 0.8)
1 0.8 Current (mA)

Erroneous data point Maximum current value ≈ 0.8 mA

0.8 Current (mA)

0.6

0.6

Maximum current value ≈ 0.3 mA
0.4

0.4

0.2

0.2

0 0 5 10 15 Time (hrs) 20 25 30

0 0 1 2 3 4 Time (Hours) 5 6 7 8

Courtesy of PNNL and KIST

Graphite felt electrode with MR-1 biofilm

Graphite felt electrode without MR-1

Graphite felt electrode with planktonic MR-1 (OD 0.4)

MR-1 biofilm/electrode images (PNNL)

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Geobiology

Many questions to answer and things to optimize However, these approaches, coupled with modeling should lead to an optimum catalyst that can be combined with optimum design to yield high power To this end: we were just awarded a MURI From DOD for this work (5 from USC). (Prakash,Ronney,Wang,Mansfeld, Nealson)

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Prospects & Approach:

Geobiology

Understand the system Optimize to produce adequate current Scale up or down for specific applications: power waste disposal remote power supplies water treatment etc.

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Geobiology
Power Production

Scale up
Waste Disposal Water Treatment Biosensors

Research tool

Same scale
Teaching

MFC

Medical Devices

Remote Power Supplies

Scale down

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Waste Disposal:

Geobiology

7 billion tons of sewage sludge generated in the US We estimate that 90% of this could be metabolized by efficient MFC approach If properly designed, we could get paid for this process by current production THANK YOU !!

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Geobiology

MICROBIAL FUEL CELLS (MFCs) Biofuels for energy production and Waste disposal THANK YOU FOR YOUR ATTENTION !! Ken Nealson Wrigley Professor of Geobiology USC (knealson@usc.edu)

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Geobiology

Microbial Fuel Cell Operation
Fuels
Organic or inorganic matter

Ion Exchange Membranes
Solid polymer or single compartment reactor

Oxidant
Atmospheric oxygen

Catalyst
Microbe at anode Pt at cathode (soon to be microbe)

-e e

e- eProton Exchange Membrane

e-

eee Anode Catalyst

eee- e e-

Cathode Catalyst

Fuel
Oxidation Reaction

e-

Oxidant
Reduction Reaction

Oxidized Fuel

H+ H+ + + H H+ H

H+ H+ + + H+H H+ H+ H

Reduced Oxidant

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Geobiology

e

e-

e- eProton Exchange Membrane

e-

eeeeOxidant eReduction Reaction

e Fuel
Oxidation Reaction

eeAnode Catalyst

Cathode Catalyst

e-

Oxidized Fuel

H+ H+
+ H+ H+ H

H+ H+
+

Reduced Oxidant

H+H

+

H+ H+ H

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