Difference between revisions of "Team:RHIT/Description"

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<h2 class="attn"> Project Description </h2>
 
<h2 class="attn"> Project Description </h2>
  
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<p>Secondary metabolites produced by microbes are often used in manufacturing processes. Due to the fact that these metabolites are produced during anaerobic respiration, being able to control, or “switch off” aerobic respiration could make industrial microbe use more efficient. <i>S. cerevisiae</i>, or baker’s yeast, is a common eukaryote used in manufacturing products including food and alcoholic beverages. This microbe is unique in that it can survive without its mitochondria by using anaerobic respiration. In order to “turn off” the mitochondria, mitochondrial ribosomal genes can be manipulated, causing mitochondrial ribosomal function to cease, therefore halting aerobic respiration. <br><br>
<span style="font-style:italic">Saccharomyces cerevisiae</span> is widely utilized in various industries in the production of alcoholic beverages, biofuel, and other organic compounds. Many of these products are secondary metabolites. Novel ways of controlling fermentation could lead to more efficient and effective methods for the industrial production of these products.  
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We created a system that allowed for control of the mitochondrial ribosomal protein S12 (<i>mRPS12</i>), using a copper repressible promoter (CTR1). In high concentrations of copper ions, this gene will be repressed and aerobic respiration will be turned off. However, copper chelator bathocuproine disulfonate (BCS), which has higher affinity for copper ions, can then be added to the system, therefore derepressing the gene and turning the mitochondria back on.<br><br>
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This concept has many applications in industry. Yeast growth follows an S-curve, with secondary metabolite generation occurring at the end of the exponential phase into the stationary phase. By regulating aerobic respiration, these yeast can begin generating secondary metabolites sooner and therefore can produce more over their lifespan. Also, due to the fact that the mitochondria would not be functioning, it is not necessary to grow these yeast in an oxygen-free environment. <br><br>
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We show that using a control system can allow for a mitochondrial “switch” that can force yeast into anaerobic respiration while in the presence of oxygen.</p>
 
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With manufacturing applications in mind, our team aims to affect production of yeast secondary metabolites through the regulation of aerobic respiration. Our project accomplishes this task by disabling translation in the mitochondria, which necessitates fermentation. Specifically, we aim to control the expression of the mitochondrial ribosomal protein S12 gene <span style="font-style:italic">(MRPS12) </span>. This protein is essential for mitochondrial translation. To control this gene, the copper sensitive promoter, CTR1, will be utilized. This system should allow us to turn aerobic respiration on and off by manipulating copper levels in the medium.
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<h2 class="attn">Previous Project Improvements</h2>
 
<h2 class="attn">Previous Project Improvements</h2>

Revision as of 16:07, 15 September 2015

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Project Description

Secondary metabolites produced by microbes are often used in manufacturing processes. Due to the fact that these metabolites are produced during anaerobic respiration, being able to control, or “switch off” aerobic respiration could make industrial microbe use more efficient. S. cerevisiae, or baker’s yeast, is a common eukaryote used in manufacturing products including food and alcoholic beverages. This microbe is unique in that it can survive without its mitochondria by using anaerobic respiration. In order to “turn off” the mitochondria, mitochondrial ribosomal genes can be manipulated, causing mitochondrial ribosomal function to cease, therefore halting aerobic respiration.

We created a system that allowed for control of the mitochondrial ribosomal protein S12 (mRPS12), using a copper repressible promoter (CTR1). In high concentrations of copper ions, this gene will be repressed and aerobic respiration will be turned off. However, copper chelator bathocuproine disulfonate (BCS), which has higher affinity for copper ions, can then be added to the system, therefore derepressing the gene and turning the mitochondria back on.

This concept has many applications in industry. Yeast growth follows an S-curve, with secondary metabolite generation occurring at the end of the exponential phase into the stationary phase. By regulating aerobic respiration, these yeast can begin generating secondary metabolites sooner and therefore can produce more over their lifespan. Also, due to the fact that the mitochondria would not be functioning, it is not necessary to grow these yeast in an oxygen-free environment.

We show that using a control system can allow for a mitochondrial “switch” that can force yeast into anaerobic respiration while in the presence of oxygen.

Previous Project Improvements

We improved the documentation of the GPD promoter previously inserted into the iGEM database. This GPD promoter had a number of different sequences with no way to differentiate between them. They had been referred to in a number of projects since creation and used interchangeably, even though they were not. Our work consisted of sorting the different GPD promoters and sequences and organizing them in a more useful manner.