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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+. | + | 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 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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| | <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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| − | 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.
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| − | How? Method ?
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| − | Lactate detection kit was used as described in “Protocol: Lactate Detection” and the following measurement were recovered:
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| | <!-- Uncomment this to enable Functional Parameter display | | <!-- Uncomment this to enable Functional Parameter display |
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color:#000000;font-weight:700;font-size:36pt;padding-bottom:6pt;font-family:"Calibri";line-height:1.0791666666666666;page-break-after:avoid;orphans:2;widows:2;text-align:left}. subtitle{ padding-top: 18pt; color:#666666;font- size:24pt;padding-bottom:4pt;font-family:"Georgia";line-height:1.0791666666666666;page-break-after:avoid;font-style:italic;orphans:2;widows:2;text-align:left }li{color: #000000; font-size:11pt;font-family:"Calibri"}p{margin :0;color:#000000;font- size: 11pt; font-family:"Calibri"}h1{padding-top:24pt;color:#000000;font-weight:700;font-size:24pt;padding-bottom:6pt;font-family:"Calibri";line-height:1.0791666666666666;page-break-after:avoid;orphans:2;widows:2;text-align:left}h2{padding-top:18pt;color:#000000;font-weight:700;font-size:18pt;padding-bottom:4pt;font-family:"Calibri";line-height:1.0791666666666666;page-break-after:avoid;orphans:2;widows:2;text-align:left}h3{padding-top:14pt;color:#000000;font-weight:700;font-size:14pt;padding-bottom:4pt;font-family:"Calibri";line-height:1.0791666666666666;page-break-after:avoid;orphans:2;widows:2;text-align:left}h4{padding-top:12pt;color:#000000;font-weight:700;font-size:12pt;padding-bottom:2pt;font-family:"Calibri";line-height:1.0791666666666666;page-break-after:avoid;orphans:2;widows:2;text-align:left}h5{padding-top:11pt;color:#000000;font-weight:700;font-size:11pt;padding-bottom:2pt;font-family:"Calibri";line-height:1.0791666666666666;page-break-after:avoid;orphans:2;widows:2;text-align:left}h6{padding-top:10pt;color:#000000;font-weight:700;font-size:10pt;padding-bottom:2pt;font-family:"Calibri";line-height:1.0791666666666666;page-break-after:avoid;orphans:2;widows:2;text-align:left}</style>< /head><body 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> | + | <html> |
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| | + | <h1>Comparison of lactate secretion between engineered and non-engineered bacteria</h1> |
| | + | <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> |
| | + | <p><img src="https://static.igem.org/mediawiki/parts/1/1d/T--Ionis_Paris--lactate-1.png" alt="Lactate concentration measurements table" /></p> |
| | + | <p><img src="https://static.igem.org/mediawiki/parts/8/84/T--Ionis_Paris--lactate-image3.png" alt="Lactate contentration chart" /> |
| | + | <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" /> |
| | + | <em>Figure 2: Evolution of lactate concentration ratio between control and modified bacteria</em></p> |
| | + | <p>These data shows clearly the efficiency of the modified organism to secrete more lactate.</p> |
| | + | </div> |
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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+.
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.
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.
