BIOB 161: PRINCIPLES OF LIVING SYSTEMS

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790#:
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Microbial Fuel Cells

Introduction/Background:
Alternative Energy and Microbial Fuel Cells
Recent rising energy costs, rapidly dwindling crude oil supplies and
concern over the negative effects of carbon emissions have reignited both
public and private interest in finding cheap alternative renewable energy
sources. Many green energy generating process rely on the metabolic
activity of microbes to turn human waste products into usable energy. For
example, you have probably heard of bioethanol as an alternative or
additive to gasoline. Ethanol is a normal by-product of the metabolism of
many microbes (e.g. in wine or beer) and can be
generated in large amounts by growing ethanolproducing microorganisms on fermentable carbon
sources like corn or grass.
Microbes can also be used to generate
electricity. As we learned in class, the energy
released during the transfer of electrons from a
donor to an acceptor when cells catabolize energy
rich molecules such as glucose can be used to
produce ATP. Electricity is really just a stream of
electrons, so, if we can find a way to steal the
electrons that microbes use in these oxidationreduction (redox) reactions, we can generate an
electric current that can be used to power electronic
devices! Microbial fuel cells (MFCs) dont have to be
complicated: any kind of chamber that provides food for microbes and a way
to steal electrons from their microbial metabolism to generate a current is
called a MFC.
Figure 1. Fosters pilot microbial fuel
cell, Queensland, Australia.
http://www.microbialfuelcell.org/www/ind
ex.php/Applications/MFC-Pilot.html

BIOB 161: PRINCIPLES OF LIVING SYSTEMS

Name:
790#:
Lab section:

History of Microbial Fuel Cells

The first microbial fuel cell was developed in 1911 (1). Throughout the
th
20 century, scientists continued to develop and test MFCs, but cheap oil
and disappointingly small electrical output from the fuel cells prevented
them from becoming a viable alternative energy source. In the last 5 years,
interest in MFCs has increased dramatically, and many businesses,
researchers and private citizens are coming up with new and better ways to
use microbes to generate electricity. In 2007, Fosters brewing company
partnered with the University of Queensland and the Australian government
to produce the worlds first operational pilot MFC that uses brewery waste as
a food source for microbes (ref. 2, Figure 1). This MFC not only generates
enough electricity to power a single-family home, it also purifies the brewery
waste water and creates a useful industrial by-product: sodium hydroxide, or
lye (3)! Think of all the soap you could make
Microbial Fuel Cells and Batteries
To understand how a MFC works, you should first understand how a
simple battery works. In a battery there are two electrodes: the anode (-)
and the cathode (+). The anode is located on one side of a chamber and
donates or gives up electrons to the cathode which is located on the
opposite side of the chamber. Using the terminology we learned in class to
describe redox reactions, the anode becomes oxidized and the cathode
becomes reduced. When you hook up an electronic device to the battery
and connect the two electrodes, electrons produced at the anode will flow to
the cathode, and the result is an electric current (Figure 2).

BIOB 161: PRINCIPLES OF LIVING SYSTEMS

Name:
790#:
Lab section:
Similarly, certain enzymes in glycolysis, for
example, reduce NAD+ to NADH with electrons
extracted from molecules in the glycolysis
pathway. In the presence of oxygen, NADH
shuttles these electrons to the electron transport
chain, ultimately reducing oxygen, the terminal
electron acceptor. Making a microbial fuel cell can
be as easy as putting microbes in a chamber with
food, letting them harvest the electrons from it for
you, and then stealing the electrons with an anode
and sending the electrons to a cathode to
generate a current.

Procedure
Today you will be working in groups of 2 or 3 to make microbial fuel
cells using bakers yeast (Saccharomyces cerevisiae), jello and carbon paper
electrodes (which conduct electricity). The purpose of this experiment is to
demonstrate that the process of catabolism generates electrons and that
these electrons can be harnessed to do work (that is, the chemical energy in
food will be converted into electrical energy). You will measure how much
voltage (electrical pressure; here, the difference in electric potential energy
between the anode and cathode) you can generate from your fuel cells.
Please read the questions beforehand and think about how you are going to
answer them as you do the lab.

BIOB 161: PRINCIPLES OF LIVING SYSTEMS

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790#:
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To make the fuel cells, you will mix together jello and yeast and put a
carbon paper electrode on either side to form a jello sandwich. The yeast
will metabolize sugar in the jello by the glycolysis pathway and generate
electrons. The top of the jello sandwich will be more exposed to air (and,
thus, oxygen) than the bottom and will therefore support a higher rate of
aerobic respiration; consequently, electrons from NADH produced in
glycolysis and the citric acid cycle will tend to reduce oxygen rather than the
carbon paper electrode. By contrast, electrons from NADH produced near the
bottom of the sandwich will reduce the electrode. For this reason, the
electrode at the top of the sandwich will be the cathode, and the bottom
electrode will be the anode. When you complete the circuit using a digital
multimeter (a device that electricians use), the electrons will move from
anode to cathode. You will be able to see how much voltage is generated on
the multimeter screen. You will make two fuel cells: one that is normal and
another that contains a chemical that inhibits glycolysis (sodium fluoride, or
NaF).
Materials
- Three 50 mL plastic tubes with caps
- Two aluminum muffin cups
- 1 plastic knife
- 80 mL jello mix solution
- teaspoon bakers yeast
- gloves
- 2 plastic droppers
- 1 thermometer
- Four squares of carbon paper measuring 1.5 in x 1.5 in.
4

BIOB 161: PRINCIPLES OF LIVING SYSTEMS

Name:
790#:
Lab section:
-

1 digital multimeter
NaF- CAUTION NaF inhibits glycolysis, which means that it
is a poison! You must wear gloves at all times, be very
careful with it and be sure to properly dispose of all the
materials that have contacted it.

Directions
1. Put on your gloves.
2. Get three 50mL plastic tubes and a styrofoam holder from the prep
area. Label one tube NaF, the second plain jello and the third
yeast.
3. Pour 40 milliliters (mL) of jello mix into the plain jello and NaF
tubes.
4. When the jello gets to 30C, measure teaspoon of dried yeast into
the yeast tube and add lukewarm tap water up to the 5 mL mark.
Let the yeast re-hydrate for a few minutes.
5. Using a plastic dropper, add 2.5 mL of yeast suspension to each jello
tube. Be sure to distribute the yeast suspension evenly to both tubes.
The yeast may settle and need to be mixed with the dropper before
dividing then between the jello tubes. Cap the tubes tightly and gently
invert the tube to mix. Please be aware that the tube caps can be
somewhat tricky to seal.
6. Using a second clean plastic dropper, carefully measure 1.5 mL of NaF
into the jello tube labelled NaF. Put your used NaF dropper in the
NaF waste bucket.
7. Seal the NaF tube. After making absolutely certain that the tube is
capped properly, gently invert the NaF tube to mix.
8. Get two aluminum muffin cups from the prep area. Label the outside
of one of the cups with your group name and plain jello and the other
with your group name and NaF.
9. Take the cups and your jello tubes to the metal tray your TA has set up
in the prep area. Place the muffin cups on the tray, give each of your
jello tubes a final mix and pour the contents of each tube into the
appropriate muffin cup.
5

BIOB 161: PRINCIPLES OF LIVING SYSTEMS

Name:
790#:
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10.

Carefully place the tray in the refrigerator until the jello is set
(~25 to 30 minutes). In the meantime, get 2 plastic plates, 4 carbon
paper squares and a digital multimeter.
11.
Once the jello is solid, remove the jello cakes from the muffin
cups and place each one on top of a carbon paper square in the middle
of a plastic plate. The carbon (black) side of the paper should be
facing up into the jello. This is the anode. To remove the jello/yeast
cakes from the muffin cups, gently run a plastic knife around the inside
edge of the cup to separate the jello from the aluminum and then tear
away the sides of the cup. Separate each jello cake from the bottom of
the muffin cup using the knife.
12.
Next, place a second carbon paper square on top of the jello cake
with the black side facing down. This is the cathode. You should have
2 jello/yeast microbial fuel cells-one with NaF and one without.
13.
Now you are ready to measure how much electricity is being
generated by your fuel cells. Turn the dial on the digital multimeter to
the millivolts setting (DCV, 200m in black lettering on the left side of
the dial).
14.
Remove the plastic caps from the ends of the multimeter leads.
Position one lead so that it makes contact with the black side of carbon
paper underneath the jello (the anode) and the other so that it makes
contact with the black side of the carbon paper on top of the jello (the
cathode). It may require two people to make sure that the leads are
simultaneously in contact with both pieces of carbon paper.
15.
Record the maximum millivolts generated by each of the fuel
cells. Also record any unusual behavior you notice. Does the mV
reading stay the same or fluctuate? Does it go up? Does it go down?
What happens if you remove the multimeter leads from the jello, allow
the fuel cell to sit for a few minutes and take another reading?
16.
Try combining your fuel cells with those from other groups. Do
more fuel cells stacked together generate more electricity?

BIOB 161: PRINCIPLES OF LIVING SYSTEMS

Name:
790#:
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Results (record the absolute values of your observations in the

table below, 20 pts)

Source

mV

Discussion (70 pts total, 10 pts

each)

1. What is the source of

electrons (i.e., the carbon
source) for the yeast in your
fuel cell? Draw this molecule
below.

2. Label the component parts (that is, the monomers of which it

is composed) of the above molecule.

3. What is the maximum millivolt reading that you measured

for each of your fuel cells?

4. How does NaF inhibit glycolysis? Be specific (use an

internet search).

BIOB 161: PRINCIPLES OF LIVING SYSTEMS

Name:
790#:
Lab section:

5. Given what you know about NaF, explain the results you got
for question 3. Why would NaF affect the amount of
electricity generated by the yeast?

6. Would your normal fuel cell continue to generate electricity

forever? Why or why not?

7. Suggest one thing that you could do to your fuel cell to

increase the voltage.

References:

1. Potter, M. C. (1911). Electrical effects accompanying the decomposition

of organic compounds. Royal Society (Formerly Proceedings of the
Royal Society) B, 84, p260-276
2. Microbial fuel-cell squeezes energy from brewery wastewater. The
New York Times, May 7, 2007.
3. New technology turns waste streams into potential income streams.
Publication, the Australian Department of Environment and Resource
Management. http://www.derm.qld.gov.au/register/p03035aa.pdf