Using Cyanobacteria to Increase Yield of Ethanol in Plants

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


Stage 2


Stage 2


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


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


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.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://biofuelsandthepoor.com/facts-and-definitions/


http://www.icgeb.org/yazdani-shams-lab.html

http://www.triplepundit.com/2012/01/doe-lab-synthetic-biology-produce-cheaper-biofuel/

http://faculty.mville.edu/ParikhS/courses/chm3049/articles/Defossiling%20Fuel%20-%20How%20Synthetic%20Biology%20Can%20Transform%20Biofuel%20Production.pdf


















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Using Cyanobacteria to Increase Yield of Ethanol in Plants