Difference between revisions of "Part:BBa K4169029"

Revision as of 07:24, 12 October 2022

Mutated TMADH

After expressing, it'll produce trimethylamine dehydrogenase (TMADH (EC 1.5.99.7)). The enzyme TMADH is an iron–sulfur flavoprotein which catalyses the oxidative demethylation of trimethylamine (TMA) to dimethylamine and formaldehyde:
(CH₃)₃N + H₂O → (CH₃)₂NH + CH₂O +2H⁺ + 2e^-. But its sequence is different from wide type. Amino acid 344 is mutated from Val to Cys. The V334C mutant is designed for wiring from the protein close to the 4Fe–4S centre, but the initial electron transfer is activated on the opposite side of the protein, close to the FMN prosthetic group, by substrate (trimethylamine) binding at the active site.

Metabolic Pathway

This enzyme is a complex iron-sulfur flavoprotein that transfers electrons to the soluble flavoprotein known as electron transferring flavoprotein. It couldn't work extracellular isolated.

Figure 1.Pathways for trimethylamine metabolism in bacteria.

Protein Molecular Structures

Trimethylamine dehydrogenase (TMADH) exist as dimers.

Figure 2.Protein molecular structures of trimethylamine dehydrogenase and it's mutated site V344C.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 388
Illegal PstI site found at 183
Illegal PstI site found at 1782
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 388
Illegal PstI site found at 183
Illegal PstI site found at 1782
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 388
Illegal XhoI site found at 1717
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 388
Illegal PstI site found at 183
Illegal PstI site found at 1782
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 388
Illegal PstI site found at 183
Illegal PstI site found at 1782
Illegal AgeI site found at 879
1000
COMPATIBLE WITH RFC[1000]

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 After expressing, it'll produce trimethylamine dehydrogenase (TMADH (EC 1.5.99.7)). The enzyme TMADH is an iron–sulfur flavoprotein which catalyses the oxidative demethylation of trimethylamine (TMA) to dimethylamine and formaldehyde: <br>(CH<sub>3</sub>)<sub>3</sub>N + H<sub>2</sub>O → (CH<sub>3</sub>)<sub>2</sub>NH + CH<sub>2</sub>O +2H<sup>+</sup> + 2e<sup>-</sup>.
+But its sequence is different from wide type. Amino acid 344 is mutated from Val to Cys. The V334C mutant is designed for wiring from  the protein close to the 4Fe–4S centre, but the initial electron  transfer is activated on the opposite side of the protein, close to  the FMN prosthetic group, by substrate (trimethylamine)  binding at the active site.
 ==Metabolic Pathway==