Difference between revisions of "Part:BBa K1587007"
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<partinfo>BBa_K1587007 short</partinfo>
 
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===BioBrick Description===
  
 
Strong RBS (BBa_1006) and formate producing enzyme pyruvate formate-lyase 1 pflB and pyruvate formate-lyase 1-activating enzyme pflA (Figure 1).
 
Strong RBS (BBa_1006) and formate producing enzyme pyruvate formate-lyase 1 pflB and pyruvate formate-lyase 1-activating enzyme pflA (Figure 1).
  
 
<center>[[File:TLSE For.jpg]]</center>
 
<center>[[File:TLSE For.jpg]]</center>
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<br>
 
<center>'''Figure 1: Genetic construction for formate production.'''</center>
 
<center>'''Figure 1: Genetic construction for formate production.'''</center>
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<br>
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===Usage and Biology===
  
 
Formate is a simple organic acid produced with an E.coli strain. The initial substrate, glucose, is decomposed into pyruvate during glycolysis, and formate is naturally synthesized thanks to two key genes:<br>
 
Formate is a simple organic acid produced with an E.coli strain. The initial substrate, glucose, is decomposed into pyruvate during glycolysis, and formate is naturally synthesized thanks to two key genes:<br>
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<center>[[File:TLSE ForMetabo.jpg]]</center>
 
<center>[[File:TLSE ForMetabo.jpg]]</center>
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<br>
 
<center>'''Figure 2: Metabolic pathway to produce formate via glucose using BBa_K1587007 in ''E. coli''.'''</center>
 
<center>'''Figure 2: Metabolic pathway to produce formate via glucose using BBa_K1587007 in ''E. coli''.'''</center>
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<br>
  
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Formate production increased significantly by 10 % compared to wt with BBa_K1587007 in ''E. coli'' (Figure 3).
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K1587007 SequenceAndFeatures</partinfo>
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===Experiments===
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In order to test BBa_K1587007 for formate production we made [http://2015.igem.org/Team:Toulouse/Experiments#erlencult micro-aerobic culture]. Samples were analyzed by [http://2015.igem.org/Team:Toulouse/Experiments#NMR NMR].
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Our main [http://2015.igem.org/Team:Toulouse/Results#formaproduct result] is shown by the graphic below:
  
 
<center>[[File:TLSE ForProd.PNG]]</center>
 
<center>[[File:TLSE ForProd.PNG]]</center>
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<br>
 
<center>'''Figure 3: NMR analysis of formate production using BBa_K1587007 after 3 days culture under micro-aerobic condition'''</center>
 
<center>'''Figure 3: NMR analysis of formate production using BBa_K1587007 after 3 days culture under micro-aerobic condition'''</center>
  
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<!-- Add more about the biology of this part here
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Formate production increased significantly by 10 % compared to wt with BBa_K1587007 in ''E. coli'' (Figure 3).
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
 
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<partinfo>BBa_K1587007 SequenceAndFeatures</partinfo>
 
  
  

Revision as of 18:09, 18 September 2015

Formate production

BioBrick Description

Strong RBS (BBa_1006) and formate producing enzyme pyruvate formate-lyase 1 pflB and pyruvate formate-lyase 1-activating enzyme pflA (Figure 1).

TLSE For.jpg


Figure 1: Genetic construction for formate production.



Usage and Biology

Formate is a simple organic acid produced with an E.coli strain. The initial substrate, glucose, is decomposed into pyruvate during glycolysis, and formate is naturally synthesized thanks to two key genes:
- pflB coding for pyruvate formate lyase which catalyzes the cleavage of pyruvate into C1 and C2. This enzyme is sensitive to oxygen and is only active in microaerobic or anaerobic conditions, which is the case within our device (Figure 2).
- pflA coding for pyruvate formate lyase activase, which is directly linked with the pyruvate formate lyase, enabling its activation (Figure 2).

TLSE ForMetabo.jpg


Figure 2: Metabolic pathway to produce formate via glucose using BBa_K1587007 in E. coli.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 405
    Illegal BamHI site found at 2807
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 514
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2995


Experiments

In order to test BBa_K1587007 for formate production we made [http://2015.igem.org/Team:Toulouse/Experiments#erlencult micro-aerobic culture]. Samples were analyzed by [http://2015.igem.org/Team:Toulouse/Experiments#NMR NMR]. Our main [http://2015.igem.org/Team:Toulouse/Results#formaproduct result] is shown by the graphic below:

TLSE ForProd.PNG


Figure 3: NMR analysis of formate production using BBa_K1587007 after 3 days culture under micro-aerobic condition

Formate production increased significantly by 10 % compared to wt with BBa_K1587007 in E. coli (Figure 3).