Team:Lethbridge/Cyanobacteria Growth

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<h2 class="pagetitle">Cyanobacteria Growth</h2>
<h2 class="pagetitle">Cyanobacteria Growth</h2>
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<p>Increasing global oil demands require new, innovative technologies for the extraction of unconventional oil sources such as those found in Alberta’s Carbonate Triangle. Carbonate oil deposits account for almost 50% of the world’s oil reserves and approximately 26% of the bitumen found in Alberta 1. Due to unstable oil prices in Western Canada, these vast reserves have historically been set aside in favour of less time consuming, more economical sites. Microbial enhanced oil recovery (MEOR) has been utilized across the world to increase the productivity of difficult resources including carbonate oil deposits. Using a synthetic biology approach, we have designed the CAB (CO2, acetic acid, and biosurfactant) extraction method that demonstrates a modified MEOR method for extracting carbonate oil deposits. CAB extraction will utilize the natural carbon fixation machinery in the cyanobacteria Synechococcus elongatus to convert CO2 into sugars to fuel acetic acid and biosurfactant production in Escherichia coli. Acetic acid applied to carbonate rock increases the pore sizes and allows for enhanced oil recovery. The reaction produces gases that will help pressurize the well site to facilitate extraction. The natural biosurfactant rhamnolipid will also be applied to the carbonate rock to further enhance extraction yields.</p>  
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<p>Establishing Growth and Handling Conditions for Synechococcus elongatus</p><br>
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<p>Overview</p>
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<p>In previous years, our team has established standard protocols and handling procedures for working with Escherichia coli. This year, we added a new organism, Synechococcus elongatus, to our repertoire and invested the necessary time for the development of detailed protocols to ensure optimal handling of this organism.</p><br>
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<p>Experimental Setup and Results</p>
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<p>We constructed a Do-It-Yourself shaking and lighting system to optimize the growth of S. elongatus (Fig. 1). S. elongatus requires light and CO2 for growth, so we monitored the growth of S. elongatus when aerated with a CO2 bubbler and without aeration. The cultures were grown at room temperature with shaking under fluorescent lighting at 400 and 700 nm. The optical density of the cultures was monitored at 750 nm for over 10 days (Fig. 2). The growth of S. elongatus progressed much faster when the culture was aerated than when it was not. This was a good indication to us that shaking does not provide enough aeration for optimal growth of S. elongatus and that using a CO2 bubbler would allow for faster growth.</p><br>
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<p><img src="https://static.igem.org/mediawiki/2012/b/b3/Cyano1.JPG"></p>
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<p>Figure 1. Shaking and lighting system used to maintain  proper growth conditions for S. elongatus. The system allows for CO2 aeration of culture flasks and direct lighting from a suspended fluorescent bulb that emits light at 400 and 700 nm.</p><br>
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<p><img src="https://static.igem.org/mediawiki/2012/a/a7/Cyano2.JPG"></p>
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<p>Figure 1. Growth curves of S. elongatus monitored by optical density at 750 nm, grown with and without aeration provided by a CO2 bubbler.</p><br>
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<p>We also tested conditions necessary for preparing glycerol stocks of S. elongatus. Glycerol stocks were prepared with 20%, 30%, and 40% glycerol and all stocks were able to be used to start cultures for growth.
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With this data, we now have working protocols that can be used to grow S. elongatus and test our constructs for enhanced glucose production and export.</p>
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<p>By coupling carbon capture with acetic acid and biosurfactant production, carbonate oil deposits can be mined with reduced greenhouse gas emissions. The use of carbon fixation to feed downstream systems can be tailored for use as a module in many applications requiring inexpensive methods for fueling biological systems. CAB extraction will be suitable for large-scale bioreactors, providing an alternative, inexpensive, and environmentally sustainable method for MEOR from Alberta’s oil deposits. Furthermore, developing the carbon capture module will be of interest in oil extraction strategies using steam, as it will help with the mitigation of CO2 release caused by steam production using for example natural gas. </p>
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Latest revision as of 03:34, 4 October 2012

2012 iGEM - University of Lethbridge

Results

Cyanobacteria Growth

Establishing Growth and Handling Conditions for Synechococcus elongatus


Overview

In previous years, our team has established standard protocols and handling procedures for working with Escherichia coli. This year, we added a new organism, Synechococcus elongatus, to our repertoire and invested the necessary time for the development of detailed protocols to ensure optimal handling of this organism.


Experimental Setup and Results

We constructed a Do-It-Yourself shaking and lighting system to optimize the growth of S. elongatus (Fig. 1). S. elongatus requires light and CO2 for growth, so we monitored the growth of S. elongatus when aerated with a CO2 bubbler and without aeration. The cultures were grown at room temperature with shaking under fluorescent lighting at 400 and 700 nm. The optical density of the cultures was monitored at 750 nm for over 10 days (Fig. 2). The growth of S. elongatus progressed much faster when the culture was aerated than when it was not. This was a good indication to us that shaking does not provide enough aeration for optimal growth of S. elongatus and that using a CO2 bubbler would allow for faster growth.


Figure 1. Shaking and lighting system used to maintain proper growth conditions for S. elongatus. The system allows for CO2 aeration of culture flasks and direct lighting from a suspended fluorescent bulb that emits light at 400 and 700 nm.


Figure 1. Growth curves of S. elongatus monitored by optical density at 750 nm, grown with and without aeration provided by a CO2 bubbler.


We also tested conditions necessary for preparing glycerol stocks of S. elongatus. Glycerol stocks were prepared with 20%, 30%, and 40% glycerol and all stocks were able to be used to start cultures for growth. With this data, we now have working protocols that can be used to grow S. elongatus and test our constructs for enhanced glucose production and export.

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