Difference between revisions of "Part:BBa K3102043"

 
 
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<partinfo>BBa_K3102043 short</partinfo>
 
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This lactate production (LDHA, LldP, ACS operon) composite part composes of Lactate dehydrogenase (LdhA):BBa_K3102026 and lactate permease (LldP):BBa_K3102027 sequences increase the production and secretion of D-Lactate in the media. The LDHA enzyme catalyses the interconversion of pyruvate and L-lactate with inter-convertion of NADH and NAD+.  
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This "lactate production" composite part is composed of Lactate dehydrogenase (ldhA):BBa_K3102026 and lactate permease (lldP):BBa_K3102027 sequences. This part increases the production and secretion of D-Lactate in the extracellular media. The LDHA enzyme catalyse the interconversion of pyruvate to L-lactate with simultaneous conversion of NADH to NAD+.
  
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This composites also constitutes of the ACS operon:BBa_K3102028, composed of three genes actP (Transports acetate) acs (catalyzes the conversion of acetate into acetyl-CoA) and yfcH (epimerase) uptake acetate molecules in order to allow the bacteria to do its metabolism and consequently survive.
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This composite part also constains the ACS operon:BBa_K3102028, composed of three genes: actP (Transports acetate), acs (catalyzes the conversion of acetate into acetyl-CoA), and yfcH (epimerase). It allows for the uptake of acetate molecules in order for the bacteria to do correctly its metabolism and survive.
  
  
 
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===Usage and Biology===
 
===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K3102043 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3102043 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K3102043 parameters</partinfo>
 
<partinfo>BBa_K3102043 parameters</partinfo>
 
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<h1>Comparison of lactate secretion between engineered and non-engineered bacteria</h1>
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<p><strong>Aims:</strong> An important step in our system in the optimization of lactate secretion by <em>E. coli</em>. Indeed, lactate constitute the energetic substrate of <em>S. oneidensis</em> and thus an essential element to the generation of electricity. Consequently, we wanted to compare lactate secretion between engineered and non-engineered bacteria.</p>
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<p><strong>Method:</strong> Lactate detection kit (Libios K-DATE Kit) was used as described in “Protocol: Lactate Detection” and the following measurement were recovered:</p>
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<p><img src="https://static.igem.org/mediawiki/parts/1/1d/T--Ionis_Paris--lactate-1.png" alt="Lactate concentration measurements table" /></p>
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<p><img src="https://static.igem.org/mediawiki/parts/8/84/T--Ionis_Paris--lactate-image3.png" alt="Lactate contentration chart" />
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<em>Figure 1: Evolution of lactate concentration between Control and the optimized E.coli</em></p>
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<p>To have a better view of the efficiency of our modified bacteria, we calculated the following ratios:</p>
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<p><img src="https://static.igem.org/mediawiki/parts/e/ec/T--Ionis_Paris--lactate-2.png" alt="Lactate ratio table" /></p>
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<p><img src="https://static.igem.org/mediawiki/parts/0/0e/T--Ionis_Paris--lactate-4.png" alt="Lactate ratio chart" />
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<em>Figure 2: Evolution of lactate concentration ratio between control and modified bacteria</em></p>
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<p>These data shows clearly the efficiency of the modified organism to secrete more lactate.</p>
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Latest revision as of 08:53, 21 October 2019


Lactate production (ldhA, lldP, ACS operon)

This "lactate production" composite part is composed of Lactate dehydrogenase (ldhA):BBa_K3102026 and lactate permease (lldP):BBa_K3102027 sequences. This part increases the production and secretion of D-Lactate in the extracellular media. The LDHA enzyme catalyse the interconversion of pyruvate to L-lactate with simultaneous conversion of NADH to NAD+.

This composite part also constains the ACS operon:BBa_K3102028, composed of three genes: actP (Transports acetate), acs (catalyzes the conversion of acetate into acetyl-CoA), and yfcH (epimerase). It allows for the uptake of acetate molecules in order for the bacteria to do correctly its metabolism and survive.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1478
    Illegal XhoI site found at 3404
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 489
    Illegal AgeI site found at 1614
    Illegal AgeI site found at 2077
    Illegal AgeI site found at 2515
    Illegal AgeI site found at 2674
    Illegal AgeI site found at 2686
    Illegal AgeI site found at 3833
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 219
    Illegal SapI.rc site found at 6698


Comparison of lactate secretion between engineered and non-engineered bacteria

Aims: An important step in our system in the optimization of lactate secretion by E. coli. Indeed, lactate constitute the energetic substrate of S. oneidensis and thus an essential element to the generation of electricity. Consequently, we wanted to compare lactate secretion between engineered and non-engineered bacteria.

Method: Lactate detection kit (Libios K-DATE Kit) was used as described in “Protocol: Lactate Detection” and the following measurement were recovered:

Lactate concentration measurements table

Lactate contentration chart Figure 1: Evolution of lactate concentration between Control and the optimized E.coli

To have a better view of the efficiency of our modified bacteria, we calculated the following ratios:

Lactate ratio table

Lactate ratio chart Figure 2: Evolution of lactate concentration ratio between control and modified bacteria

These data shows clearly the efficiency of the modified organism to secrete more lactate.