Difference between revisions of "Team:Calgary/Synthesis"

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<br><h2>Overview</h2>
 
<br><h2>Overview</h2>
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<p>Our goal was to produce polyhydroxybutyrate (PHB) by utilizing the nutrients present in human waste. Thus, the first step of our project was to design efficient and optimal pathways that can utilize the abundant and useful nutrients present in human waste to make bioplastic. After analyzing the composition of fermented synthetic human waste, we decided to focus on two major nutrients: acetyl-coA and volatile fatty acids (VFAs). Acetyl-coA is the end product of the glycolysis pathway, which is converted to acetic acid (...% in supernatant) <source>. Volatile fatty acids include ….. and are present in  …% in fermented supernatant <source>. Thus, by using products of glycolysis and volatile fatty acids in human waste, our pathway utilizes …% of the carbon source <source>. Finally, in order to convert the feedstock nutrients into PHB a naturally existing pathway in R. eutropha that converts acetyl-coA to PHB and beta-oxidation pathway that converts VFAs to PHB was selected.  
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<p>The overarching goal for the synthesis component of the project was to produce poly-3-hydroxybutyrate (PHB) by utilizing the nutrients present in human waste. In order to accomplish this, we:
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<ol>
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<li>analyzed human waste and chose organic compounds to use as feedstocks for our bacteria, and</li>
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<li>researched and optimized efficient pathways to turn relevant components of human waste (acetyl-CoA and volatile fatty acids) into PHB</li>
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</ol>
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</p>
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<h2>Chassis and vector</h2>
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    Our gene construct was placed under an inducible promoter (lacZ, lacY). The construct was inserted into pET29(b)+ vector, which contains the promoter and kan resistant gene. The chassis used for our experiments was E. coli (BL21) because of its better ability in expression of proteins (source). (Add more reasons for why e. Coli was used) The operon was induced using IPTG at different concentrations.
 
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Revision as of 00:23, 31 October 2017

Header

Synthesis

Synthesis Pathway

Overview

The overarching goal for the synthesis component of the project was to produce poly-3-hydroxybutyrate (PHB) by utilizing the nutrients present in human waste. In order to accomplish this, we:

  1. analyzed human waste and chose organic compounds to use as feedstocks for our bacteria, and
  2. researched and optimized efficient pathways to turn relevant components of human waste (acetyl-CoA and volatile fatty acids) into PHB

Chassis and vector

Our gene construct was placed under an inducible promoter (lacZ, lacY). The construct was inserted into pET29(b)+ vector, which contains the promoter and kan resistant gene. The chassis used for our experiments was E. coli (BL21) because of its better ability in expression of proteins (source). (Add more reasons for why e. Coli was used) The operon was induced using IPTG at different concentrations.

WORKS CITED

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