Difference between revisions of "Team:Dalhousie/Experiments"

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  <h1  style="color:white;"> Experiments </h1>
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<h4 style="text-align:center; color: white;">Background</h4>
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Canada’s forests represent not only an established source of economic revenue, but also a potential source for biofuel substrate. Ligno-cellulosic by-products from pulp and paper processing are removed using chemicals, heat, and water. Microbial cellulose-degrading enzymes are high-value targets for industrial applications. Focused on innovative applications of synthetic biology, the undergraduate Dalhousie iGEM team has undertaken a multi-year project to harness the degradative capacity of microorganisms to convert cellulose into ethanol for biofuel applications. Advances in DNA sequencing technology and bioinformatics have revolutionized our ability to identify useful genes in complex biological samples. We hypothesize that the porcupine microbiome, which includes microorganisms capable of digesting bark and tree resin, will be a rich source of these useful genes. If our hypothesis is correct, mining the porcupine microbiome has the advantage of finding a suite of enzymes that have evolved to work in concert to efficiently degrade cellulose. </p>
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<h4 style="text-align:center; color: white;">The Project</h4>
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Using a combination of metagenomic library construction and bioinformatic analysis we aim to find known cellulolytic enzymes as well as discover novel enzymes.
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Screening of our metagenomic library will allow us to look for a diverse selection of enzymes, including cellulolytic and lignolytic ones by plating on cellulose-only media. The clones that grow can be sequenced for confirmation and cloned into biobricks.
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Our new bioinformatic pipeline now allows us to look for conserved domains rather than known enzymes. This is extremely powerful as it may allow us to discover new enzymes that share catalytic domains with cellulases but are otherwise totally novel.
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The goal of this project is to construct a bioreactor where cellulose would be the input, D-glucose the output and E.coli expressing a suite of cellulolytic enzymes would be the workhorse, converting one to the other. Future goals would be to combine this in co-culture with yeast for the last step in bio-ethanol production.</p>
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<div class="top" ><div class="title" >Experiments</div></div>
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</br>Project Description</br>
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WHY</br>
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As fossil fuels continue to run out across the globe, many people are looking to alternate sources of energy that are renewable and eco-friendlier. One of these options is biofuel which is fuel made from organic matter. Most commonly made from ethanol, biofuel or bioethanol can be used as a fuel for vehicles in its’ pure form. In the field of biofuel production, bioethanol made from cellulose continues to be the dominant form. However harsh methods are required to be able to extract the sugars from cellulose and convert it to ethanol.
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Atlantic Canada’s main export is wood, pulp, and paper. Many of us on the team grew up next to pulp and paper mills and saw the hazardous waste expelled by the processes in these buildings. Extraction of usable materials from wood is fairly inefficient, leaving behind wood waste that could be used for biofuel production if broken down and converted to ethanol. The only question is: how?</br>
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WHAT</br>
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The Dalhousie iGEM team this year is focused on using the microbiome of the porcupine to solve this conversion of wood waste to ethanol. A huge part of the porcupine’s diet is made from bark. Unable to digest the cellulose, hemi-cellulose, and lignin in the bark, the gut bacteria of the porcupine do the work instead. We hypothesized that the microbiome of the porcupine would contain enzymes that convert cellulose, hemi-cellulose, and lignin to glucose; a usable sugar. We then hypothesized that if the genes coding for these enzymes were expressed in a vector in E. coli, the E. coli would then be able to digest cellulose and create glucose. Finally, we hypothesized that a bioreactor system containing both E. coli and yeast would be able to create ethanol from wood waste as the yeast would ferment the glucose created by the E. coli.</br>
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HOW WE DID IT</br>
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To be assessed later</br>
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Learn more... hopefully have links to next pages here</p>
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Revision as of 13:13, 4 October 2017

Experiments

Project Description
WHY
As fossil fuels continue to run out across the globe, many people are looking to alternate sources of energy that are renewable and eco-friendlier. One of these options is biofuel which is fuel made from organic matter. Most commonly made from ethanol, biofuel or bioethanol can be used as a fuel for vehicles in its’ pure form. In the field of biofuel production, bioethanol made from cellulose continues to be the dominant form. However harsh methods are required to be able to extract the sugars from cellulose and convert it to ethanol. Atlantic Canada’s main export is wood, pulp, and paper. Many of us on the team grew up next to pulp and paper mills and saw the hazardous waste expelled by the processes in these buildings. Extraction of usable materials from wood is fairly inefficient, leaving behind wood waste that could be used for biofuel production if broken down and converted to ethanol. The only question is: how?
WHAT
The Dalhousie iGEM team this year is focused on using the microbiome of the porcupine to solve this conversion of wood waste to ethanol. A huge part of the porcupine’s diet is made from bark. Unable to digest the cellulose, hemi-cellulose, and lignin in the bark, the gut bacteria of the porcupine do the work instead. We hypothesized that the microbiome of the porcupine would contain enzymes that convert cellulose, hemi-cellulose, and lignin to glucose; a usable sugar. We then hypothesized that if the genes coding for these enzymes were expressed in a vector in E. coli, the E. coli would then be able to digest cellulose and create glucose. Finally, we hypothesized that a bioreactor system containing both E. coli and yeast would be able to create ethanol from wood waste as the yeast would ferment the glucose created by the E. coli.
HOW WE DID IT
To be assessed later

Learn more... hopefully have links to next pages here