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Team:TMU-Tokyo/Project device2
From 2012.igem.org
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| - | <i>Escherichia coli</i> has original formaldehyde dehydrogenase(FDH), but it depends on glutathione existence. Instead of it, we found glutathione-independent FDH in <i>Pseudomonas putida</i>. Formaldehyde dehydrogenase changes formaldehyde to formate.(HCHO → HCOOH)<Br> | + | <i>Escherichia coli</i> has original formaldehyde dehydrogenase (FDH), but it depends on glutathione existence. Instead of it, we found glutathione-independent FDH (PFDH) in <i>Pseudomonas putida</i>. Formaldehyde dehydrogenase changes formaldehyde to formate. (HCHO → HCOOH)<Br> |
<Br> | <Br> | ||
| - | The reason we didn't select <i>Escherichia coli</i>'s own FDH is that glutathione-independent one can act in much more situations. For example, if Cheff Ant E.coli dies, <i>Escherichia coli</i>'s own FDH loses its substrate because of glutathione absence.<Br> | + | The reason we didn't select <i>Escherichia coli</i>'s own FDH is that glutathione-independent one can act in much more situations. For example, if Cheff Ant E.coli dies, <i>Escherichia coli</i>'s own FDH loses its substrate because of glutathione absence. FDH and PFDH are homologous and their sequences are well preserved.<Br> |
<Br> | <Br> | ||
| - | This device is composed of its gene and constitutive promoter.<Br> | + | In additon to the glutathione-independence, PFDH can be increased its activity by a few amino acids mutation. It was measured that the activity increased 1.7 times stronger.<Br> |
| + | <Br> | ||
| + | This device is composed of its gene and constitutive promoter. The reason we don't use repressor like <i>frmR</i> is to increase resistance of formaldehyde. We use the same strategy in <A Href="https://2012.igem.org/Team:TMU-Tokyo/Project_device3">device3</A> to increase resistance of formate.<Br> | ||
<Br> | <Br> | ||
</p> | </p> | ||
Revision as of 03:22, 14 September 2012
Device2;
Producing formaldehyde dehydrogenase, changing formaldehyde to formate
Escherichia coli has original formaldehyde dehydrogenase (FDH), but it depends on glutathione existence. Instead of it, we found glutathione-independent FDH (PFDH) in Pseudomonas putida. Formaldehyde dehydrogenase changes formaldehyde to formate. (HCHO → HCOOH)
The reason we didn't select Escherichia coli's own FDH is that glutathione-independent one can act in much more situations. For example, if Cheff Ant E.coli dies, Escherichia coli's own FDH loses its substrate because of glutathione absence. FDH and PFDH are homologous and their sequences are well preserved.
In additon to the glutathione-independence, PFDH can be increased its activity by a few amino acids mutation. It was measured that the activity increased 1.7 times stronger.
This device is composed of its gene and constitutive promoter. The reason we don't use repressor like frmR is to increase resistance of formaldehyde. We use the same strategy in device3 to increase resistance of formate.
