Difference between revisions of "Part:BBa K5059001"

 
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<partinfo>BBa_K5059001 short</partinfo>
 
<partinfo>BBa_K5059001 short</partinfo>
  
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Integrated a strong constitutive promoter (BBa_K124002), Kozak sequence (BBa_K165002), alpha-amyrin synthase (AAS) gene (BBa_K5059000), and a strong terminator (BBa_K2926005).  
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This is a composite part that integrates a strong constitutive promoter (BBa_K124002), Kozak sequence (BBa_K165002), Team Cornell's BBA_K5059000 BioBrick that encodes for alpha-amyrin synthase (AAS), and a strong terminator (BBa_K2926005). Implementing these BioBricks together allows for increased expression of AAS, outputting more alpha-amyrin for ursolic acid production.
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===Sequence and Features===
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<partinfo>BBa_K5059000 SequenceAndFeatures</partinfo>
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<!-- Uncomment this to enable Functional Parameter display
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===Functional Parameters===
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<partinfo>BBa_K5059001 parameters</partinfo>
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<!-- Add more about the biology of this part here
 
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===Usage and Biology===
 
===Usage and Biology===
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Ursolic acid has gained traction recently as a potential therapeutic agent. Preliminary studies have been done on ursolic acid, determining its therapeutic potential for cancer, liver disease, and obesity, among other health benefits [2]. There is particular interest in ursolic acid's properties in fighting cancer, as it is an antioxidant and anti-inflammatory agent. Clinical trials are currently underway to test its use in cancer-treating drugs. However, the current method for ursolic acid extraction from fruits, such as apples and loquats, is inefficient, environmentally taxing, and expensive. By engineering S. cerevisiae to produce ursolic acid, the traditional method for its extraction can be bypassed by utilizing the pathway shown below.
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K5059001 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K5059001 SequenceAndFeatures</partinfo>
  
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<html>
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<center>
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<img src="https://static.igem.wiki/teams/5059/pathway.webp" style="width: 600px; height:auto">
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<figcaption>Figure 1: Metabolic Pathway for Producing Ursolic Acid in Yeast</figcaption>
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</center>
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</html>
  
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<!-- Uncomment this to enable Functional Parameter display
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Within this pathway, TDH3-AAS-TPS1 is the first enzyme required to catalyze ursolic acid production. Specifically, it catalyzes the conversion of 2,3-oxidosqualene to alpha-amyrin, which is the direct precursor for ursolic acid through the action of Cytochrome P450, which is activated via Cytochrome P450 reductase donating an electron from NADPH to it.
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===Functional Parameters===
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<partinfo>BBa_K5059001 parameters</partinfo>
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<h2>Functionality</h2>
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===Catalytic Efficiency===
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<center>
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<table>
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  <tr>
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    <th>Substrate</th>
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    <th>Product</th>
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    <th>Km (µM)</th>
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    <th>kcat (min^-1) </th>
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    <th>kcat/Km (min^-1/µM)</th>
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  </tr>
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  <tr>
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    <td>1. 2,3-oxidosqualene</td>
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    <td>alpha-amyrin</td>
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    <td>50.07</td>
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    <td>43.4</td>
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    <td>0.87</td>
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  </tr>
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  <tr>
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    <td>2. alpha-amyrin</td>
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    <td>Ursolic Acid</td>
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    <td>24.5</td>
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    <td>35</td>
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    <td>1.43</td>
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  </tr>
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</table>
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</center>
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<center>This data comes from Dr. Yu et al.[3].</center>

Revision as of 13:41, 1 October 2024


TDH3-AAS-TPS1

This is a composite part that integrates a strong constitutive promoter (BBa_K124002), Kozak sequence (BBa_K165002), Team Cornell's BBA_K5059000 BioBrick that encodes for alpha-amyrin synthase (AAS), and a strong terminator (BBa_K2926005). Implementing these BioBricks together allows for increased expression of AAS, outputting more alpha-amyrin for ursolic acid production.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Figure 1: Metabolic Pathway for Producing Ursolic Acid in Yeast

Within this pathway, TDH3-AAS-TPS1 is the first enzyme required to catalyze ursolic acid production. Specifically, it catalyzes the conversion of 2,3-oxidosqualene to alpha-amyrin, which is the direct precursor for ursolic acid through the action of Cytochrome P450, which is activated via Cytochrome P450 reductase donating an electron from NADPH to it.

Functionality

Catalytic Efficiency

Substrate Product Km (µM) kcat (min^-1) kcat/Km (min^-1/µM)
1. 2,3-oxidosqualene alpha-amyrin 50.07 43.4 0.87
2. alpha-amyrin Ursolic Acid 24.5 35 1.43
This data comes from Dr. Yu et al.[3].