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Team:Exeter/enzyme-kinetics
From 2014.igem.org
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<td>NemA Vmax (TNT)</td> | <td>NemA Vmax (TNT)</td> | ||
<td>XenB Vmax (TNT)</td> | <td>XenB Vmax (TNT)</td> | ||
| + | <td>NemA Vmax (Nitroglycerin)</td> | ||
| + | <td>XenB Vmax (Nitroglycerin)</td> | ||
| + | <td>NemA Km (TNT)</td> | ||
| + | <td>XenB Km (TNT)</td> | ||
<td>NemA Km (Nitroglycerin)</td> | <td>NemA Km (Nitroglycerin)</td> | ||
<td>XenB Km (Nitroglycerin)</td> | <td>XenB Km (Nitroglycerin)</td> | ||
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<tr> | <tr> | ||
<td>Experimental Results</td> | <td>Experimental Results</td> | ||
| + | <td>-</td> | ||
| + | <td>-</td> | ||
| + | <td>8</td> | ||
| + | <td>-</td> | ||
| + | <td>-</td> | ||
| + | <td>-</td> | ||
| + | <td>-</td> | ||
| + | <td>15</td> | ||
</tr> | </tr> | ||
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Revision as of 12:48, 27 September 2014
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Kinetic Analysis of NemA and XenB by HPLC
Conclusion
NemA and Xen B are capable of catalysing the conversion of TNT to various products using NADH and FMN as cofactors. The binding affinity of each protein for this substrate (the Michealis Menten constant, Km) and the maximum reaction velocity (Vmax), were determined and are comparable to the published values; shown in figure 1. NemA and XenB are therefore suitable enzymes for use in our system and have been shown to function at the physiologically relevant pH of 7.
Abstract
NemA and XenB are two proteins that the iGEM Exeter team propose will allow E.coli to degrade TNT, at concentrations above those normally toxic to the cell. Among many others proposed, NemA catalyses the reaction shown in figure 2.
Figure 2 NemA Vmax (TNT) XenB Vmax (TNT) NemA Vmax (Nitroglycerin) XenB Vmax (Nitroglycerin) NemA Km (TNT) XenB Km (TNT) NemA Km (Nitroglycerin) XenB Km (Nitroglycerin) Experimental Results - - 8 - - - - 15 Published Values
Exeter | ERASE
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