Using Cyanobacteria to Increase
Yield of Ethanol in Plants
By: Vishal Mummareddy
Ethanol Production in
the U.S.
Americans used 13 billion gallons of Ethanol last year
World Ethanol production has tripled from 2000 to
2007
36 billion gallons of Ethanol will be produced during
the year 2022
How Ethanol is Made?
Cellulose and
Hemicellulose
Cellulose and Hemicellulose are the sugars used to make
ethanol
They are found in the cell wall of most plant cells
Lignin is intertwined with cellulose and hemicellulose and
must be seperated in order to make ethanol
Cellulose and Hemicellulose are made of thousands of
glucose units
So if we can increase glucose production, we can increase
cellulose production which will then create more ethanol
Cellulose and
Hemicellulose
Plant Cells
Many plant pathogens infect plants through the
roots, but some of them can get in through the
stomata
The stomata is used for gas exhange and opens when
there are changes in the environment
The purpose of these bacteria are to colonize the
plant and eventualy kill the plant
Pseudomonas syringae is a bacterial plant pathogen
that can go through the stomata of plant leaves
Pseudomonas syringae
Is an airborne bacteria that infects many different
types of plants
It causes the water in plants to freeze which results
in injury for the plants
Cyanobacteria
Cyanobacteria is one of the most productive
photosynthetic bacteriums
This type of bacteria can be found in almost every
terrestial and aquatic habitat
For my design project, I am going to use
cyanobacteria and the components of Pseudomonas
syringae to infultrate plant leaves and increase
glucose production
Getting Cyanobacteria
into Plants
The leaf of plants is a great place for bacteria to grow
and replicate because: 1. Moist 2. Sunlight
In order for cyanobacteria to invade the plant it must
release virulence effector proteins into the plant
Getting Cyanobacteria
into Plants
To trigger the proteins, the farmer must spray
Gibberellic Acid: a growth hormone for plants
This hormone will then release the virulence effector
proteins in the cyanobacteria which will then trick
the plant into letting in the cyanobacteria
These proteins can be found in Pseudomonas syringae
Then the bacteria will grow into the plant and travel
to the spongy mesophyl
Plant Leaf
Cyanobacteria Entering
Plant: Stage 1
Cyanobacteria Entering
Plant: Stage 2
Cyanobacteria Entering
Plant: Stage 2
Cyanobacteria Entering
Plant: Stage 3
Cyanobacteria in Plant
Leaf
The cyanobacteria will connect to the plant cell using the hrpgene cluster.
This gene contains a needle that extends from the
cyanobacteria
This needle is used to transport proteins that will infect the host
cell
However, once all the proteins are taken out, it can be used to
export glucose
Size of protein - 3 – 6 nm
Size of glucose molecule – 1 nm
Cyanobacteria Joining
with Plant Cell
Cyanobacteria Joining
with Plant Cell
Engineering
Cyanobacteria
The first step of the proccess would be to get the
bacteria into the plant
To get this done, cyanobacteria needs to secrete
virulence effector proteins
The effector protein tricks the stomata into letting
the bacteria inside
Engineering
Cyanobacteria
In addition to the virulence effector proteins, the
cyanobacteria needs a hrp-gene cluster
This gene cluster contains the coding for a base,
needle, and tip wich can be used to transfer
molecules into the host cell
Engineering
Cyanobacteria
Expected Results
Conjoined with Plant Cell
Transfer Glucose
0
1
0
1
The bacteria should release glucose using the
chaperone molecules when it is conjoined with the
plant cell
Making Ethanol
Now that we have a bacteria that can make and
transport glucose into the plant, we can then
increase the yield of ethanol
This happens because the plant cells store the extra
glucose in forms of starch in the leaves, roots, or
stems
Making Ethanol
After the crops are harvested, it undergoes 5 major
processes in order to make ethanol
1.
2.
3.
4.
5.
Milling
Liquefaction
Saccharification
Fermentation
Distillation and Recovery
Milling
This is when the crop is smashed in order to get it
into its basic components
Liquefaction
Is when the smashed crop is heated and the enzymes
break down the starches into smaller fragments
Saccharification
The starches are then broken down into glucose
Fermentation
Yeast is then added to the glucose to start the
process of converting glucose into ethanol
Distillation and
Recovery
The ethanol is then removed and used for fuel
Advantages
My design will always be able to increase glucose
production because most companies focus on
making the plant more productive. But my design
will always work and act as and additive for glucose
production
Advantages
This design will help increase the yield of ethanol
production in plants. This will definitely make
ethanol more economical. As a result, more
consumers will start to use ethanol.
Potential Problems
Some bacteria may not be able to enter the plant or
may die in the mesophyll
The bacteria could mutate which can lead to the
bacterium harming the plant
The bacteria could travel and infect other plants and
harm other crops
Testing
I would test the bacteria on plants in a lab setting and
measure the production of ethanol from a plant with
my bacteria and one without it. This will prove that
my bacterium is effective and can increase the yield
of ethanol in plants.
Works Cited
http://www.ncbi.nlm.nih.gov/pubmed/18077204
http://www.unl.edu/alfanolab/pdf/publications/14.pdf
http://en.wikipedia.org/wiki/Pseudomonas_syringae
http://www.pnas.org/content/97/16/8770.abstract
http://www.sciencedirect.com/science/article/pii/S0092867406010154
http://www.ncbi.nlm.nih.gov/pubmed/17419713
http://www.ncbi.nlm.nih.gov/pubmed/16439654
http://www.extension.purdue.edu/extmedia/ID/ID-328.pdf
Works Cited
http://faculty.mville.edu/ParikhS/courses/chm3049/articles/Defossiling%20F
uel%20%20How%20Synthetic%20Biology%20Can%20Transform%20Biofuel%20Produc
tion.pdf
http://www.epa.gov/ttnecas1/regdata/IPs/Starch%20Manufacturing_IP.pdf
http://www.ncbi.nlm.nih.gov/pubmed/18476862
http://biofuelsandthepoor.com/facts-and-definitions/
http://www.sciencedirect.com/science/article/pii/S0958166908001420
http://www.icgeb.org/yazdani-shams-lab.html
http://www.triplepundit.com/2012/01/doe-lab-synthetic-biology-producecheaper-biofuel/