Part:BBa_K4169029

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.[1]

Metabolic Pathway

This enzyme is a complex iron-sulfur flavoprotein that transfers electrons to the soluble flavoprotein known as electron transferring flavoprotein.[2] It couldn't work extracellular isolated.

Figure 1.Pathways for trimethylamine metabolism in bacteria.

Protein Molecular Structures

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

Sequence and Features

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]