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Using Cyanobacteria to Increase Yield of Ethanol
in PlantsBy: 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 CellsMany 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
CyanobacteriaCyanobacteria 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 PlantsThe 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 PlantsTo 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 hrp-gene 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
The bacteria should release glucose using the chaperone molecules when it is conjoined with the plant cell
Conjoined with Plant Cell
Transfer Glucose
0 01 1
Making EthanolNow 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 EthanolAfter the crops are harvested, it undergoes 5 major processes in order to make ethanol1. Milling
2. Liquefaction
3. Saccharification
4. Fermentation
5. Distillation and Recovery
Milling This is when the crop is smashed in order to get it into its basic components
LiquefactionIs when the smashed crop is heated and the enzymes break down the starches into smaller fragments
SaccharificationThe starches are then broken down into glucose
FermentationYeast 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 Citedhttp://faculty.mville.edu/ParikhS/courses/chm3049/articles/Defossiling%20Fuel%20-%20How%20Synthetic%20Biology%20Can%20Transform%20Biofuel%20Production.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-produce-cheaper-biofuel/