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 "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.
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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class="c20"><p class="c8"><span class="c7">Results: Comparison of lactate secretion between engineered and non-engineered bacteria </span></p><p class="c8 c22"><span class="c7"></span></p><p class="c9"><span>An important step in our system in the optimization of lactate secretion by </span><span class="c11">E. coli</span><span>. Indeed, lactate constitute the energetic substrate of </span><span class="c11">S. oneidensis</span><span class="c4"> and thus an essential element to the generation of electricity. </span></p><p class="c9"><span class="c4">Lactate detection kit was used as described in “Protocol: Lactate Detection” and the following measurement were recovered:</span></p><p class="c9 c22"><span class="c4"></span></p><a id="t.c0c51514b21a288cf077105e6ffccb79c8a44457"></a><a id="t.0"></a><table class="c18"><tbody><tr class="c3"><td class="c5" colspan="1" rowspan="1"><p class="c2"><span class="c0"> </span></p></td><td class="c5" colspan="1" rowspan="1"><p class="c2"><span class="c0">0h</span></p></td><td class="c5" colspan="1" rowspan="1"><p class="c2"><span class="c0">3h</span></p></td><td class="c5" colspan="1" rowspan="1"><p class="c2"><span class="c0">6h</span></p></td><td class="c5" colspan="1" rowspan="1"><p class="c2"><span class="c0">9h</span></p></td></tr><tr class="c14"><td class="c6 c16" colspan="1" rowspan="1"><p class="c2"><span class="c12">Control </span></p></td><td class="c6" colspan="1" rowspan="1"><p class="c2"><span class="c4">60,235</span></p></td><td class="c6" colspan="1" rowspan="1"><p class="c2"><span class="c4">49,982</span></p></td><td class="c6" colspan="1" rowspan="1"><p class="c2"><span class="c4">24,671</span></p></td><td class="c6" colspan="1" rowspan="1"><p class="c2"><span class="c4">13,136</span></p></td></tr><tr class="c14"><td class="c6 c16" colspan="1" rowspan="1"><p class="c2"><span class="c12">LLOpLdha </span></p></td><td class="c6" colspan="1" rowspan="1"><p class="c2"><span class="c4">69,206</span></p></td><td class="c6" colspan="1" rowspan="1"><p class="c2"><span class="c4">67,925</span></p></td><td class="c6" colspan="1" rowspan="1"><p class="c2"><span class="c4">47,740</span></p></td><td class="c6" colspan="1" rowspan="1"><p class="c2"><span class="c15">48,060</span></p></td></tr></tbody></table><p class="c8"><span style="overflow: hidden; display: inline-block; margin: 0.00px 0.00px; border: 0.00px solid #000000; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px); width: 514.40px; height: 314.40px;"><img alt="" src="images/image1.png" style="width: 514.40px; height: 314.40px; margin-left: 0.00px; margin-top: 0.00px; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px);" title=""></span></p><p class="c21"><span class="c4">To have a better view of the efficiency of our modified bacteria, we calculated the following ratios:</span></p><p class="c10"><span class="c4"></span></p><a id="t.fcbaf46168b43ac8baa366cf42d318313bd37c92"></a><a id="t.1"></a><table class="c18"><tbody><tr class="c17"><td class="c1" colspan="1" rowspan="1"><p class="c2"><span class="c0"> </span></p></td><td class="c1" colspan="1" rowspan="1"><p class="c2"><span class="c0">0h</span></p></td><td class="c1" colspan="1" rowspan="1"><p class="c2"><span class="c0">3h</span></p></td><td class="c1" colspan="1" rowspan="1"><p class="c2"><span class="c0">6h</span></p></td><td class="c1" colspan="1" rowspan="1"><p class="c2"><span class="c0">9h</span></p></td></tr><tr class="c13"><td class="c1" colspan="1" rowspan="1"><p class="c2"><span class="c12">Ratio</span></p></td><td class="c19" colspan="1" rowspan="1"><p class="c2"><span class="c0">1,15</span></p></td><td class="c19" colspan="1" rowspan="1"><p class="c2"><span class="c0">1,36</span></p></td><td class="c19" colspan="1" rowspan="1"><p class="c2"><span class="c0">1,93</span></p></td><td class="c19" colspan="1" rowspan="1"><p class="c2"><span class="c0">3,66</span></p></td></tr></tbody></table><p class="c8"><span style="overflow: hidden; display: inline-block; margin: 0.00px 0.00px; border: 0.00px solid #000000; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px); width: 457.60px; height: 271.20px;"><img alt="" src="images/image2.png" style="width: 457.60px; height: 271.20px; margin-left: 0.00px; margin-top: 0.00px; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px);" title=""></span></p><p class="c21" id="h.gjdgxs"><span class="c4">This data shows clearly the efficiency of the modified organism to secrete more lactate. </span></p></html>
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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>
+
<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>
<em>Figure 1: Evolution of lactate concentration between Control and the optimized E.coli</em></p>
+
<p>To have a better view of the efficiency of our modified bacteria, we calculated the following ratios:</p>
+
<p><img src="https://static.igem.org/mediawiki/parts/e/ec/T--Ionis_Paris--lactate-2.png" alt="Lactate ratio table" /></p>
+
<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:
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:
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.
Figure 1: Evolution of lactate concentration between Control and the optimized E.coli
Figure 2: Evolution of lactate concentration ratio between control and modified bacteria