Team:BV CAPS Kansas/Project

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<h1> Project </h1>
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<p>Project Overview</p>
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<p>Abstract - The Effects of Increased Pyruvate Kinase Expression on the Production of Alkanes in Cyanobacteria</p>
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Alkanes, the primary component in diesel, are one class of molecules that cyanobacteria can synthesize. Furthermore, alkanes excreted from the cell then float on the surface of water, making harvesting of alkanes efficient and fairly simple through dewatering. Like other fatty-acid-derived products, acetyl-CoA is the initiation point for synthesis of alkanes. In a photosynthetic system such as cyanobacteria, acetyl-CoA is formed from pyruvate, which is the end product of glycolysis. Since pyruvate kinase catalyzes the final step in glycolysis (the conversion of phosphoenolpyruvate (PEP) to pyruvate), this enzyme is a common regulatory point of glycolysis in most organisms. Because the production of pyruvate provides the initial carbon source for lipid and alkane synthesis, we hypothesize that increasing the enzymatic activity of pyruvate kinase will increase the cellular concentration of pyruvate and thus increase the production of alkanes by the cell.
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Humans rely on carbon resources for nutrition and energy. Through industrialization we have become dependent on non-renewable fossil fuels to the detriment of the environment. Recent research into renewable biofuels has included work on the production of corn-based ethanol and the microbial degradation of cellulose from terrestrial plants. Biofuels derived from these sources are not fully sustainable. Third generation biofuels include those derived from microalgae. Cyanbacteria are microalgae known to fix carbon dioxide into alkanes through the collective processes of photosynthesis, glycolysis, and fatty acid biosynthesis. The CAPS iGEM Team 2013 begins metabolic engineering of these pathways by expressing a rabbit muscle derived pyruvate kinase, known to be a key regulator of glycolysis, within the cyanobacteria Synechocystis PCC 6803 in an effort to increase alkane production. Pathways will be modeled using TinkerCell and assays for the production of pyruvate, fatty acids, and alkanes will be used to characterize our system.
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In order to increase alkane production, we will insert a more activate pyruvate kinase gene in cyanobacteria (Synechocystis PCC 6803) in a broad host range plasmid (part BBa_K12500). The system could then be measured for pyruvate and alkanes through gas chromatography and a lipid profiling assay.  
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Revision as of 14:05, 21 June 2013

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Project

Abstract - The Effects of Increased Pyruvate Kinase Expression on the Production of Alkanes in Cyanobacteria

Humans rely on carbon resources for nutrition and energy. Through industrialization we have become dependent on non-renewable fossil fuels to the detriment of the environment. Recent research into renewable biofuels has included work on the production of corn-based ethanol and the microbial degradation of cellulose from terrestrial plants. Biofuels derived from these sources are not fully sustainable. Third generation biofuels include those derived from microalgae. Cyanbacteria are microalgae known to fix carbon dioxide into alkanes through the collective processes of photosynthesis, glycolysis, and fatty acid biosynthesis. The CAPS iGEM Team 2013 begins metabolic engineering of these pathways by expressing a rabbit muscle derived pyruvate kinase, known to be a key regulator of glycolysis, within the cyanobacteria Synechocystis PCC 6803 in an effort to increase alkane production. Pathways will be modeled using TinkerCell and assays for the production of pyruvate, fatty acids, and alkanes will be used to characterize our system.

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