Part:BBa_K4680004

R. pomeroyi TMM Generator

Usage and Biology

The FSU iGEM team created this biological device to oxide trimethylamine (TMA) in the small intestine of a human. The purpose behind this device is to eliminate TMA from the body for those struggling with trimethylaminuria (TMAU). TMAU is a rare condition (or disease) in which a mutation appears on the FMO3 enzyme coding sequence and the body can no longer oxidize trimethylamine into trimethylamine N-oxide in the liver. This causes the person to expel a fishy odor throughout their entire body. Our device consists of an Anderson promoter, synthetic RBS, a TMM enzyme coding sequence taken from the Paris Bettencourt 2014 team, and a terminator. The trimethylamine monooxygenase coding sequence comes from a species called Ruegeria Pomeroyi. This species comes from the Marine Roseobacter clade that uses TMAO as a nutrient source. The Vmax and Km values of the TMM enzyme are 15.8 +/- 3.1 μg*min-1*mg-1 and 20.8 +/- 2.9 μM. These values are higher than the human FMO3 enzyme values, therefore we assume it would work better than the human FMO3 enzyme.

Measurements

This biological device has been inserted in a pIDTSmart-Kan plasmid backbone from IDT resulting in pFSUIGEM201. Figure 1 depicts the map of pFSUIGEM201.

Figure 1 - pFSUIGEM201

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 432
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 256
  Illegal AgeI site found at 479
  
• 1000
  COMPATIBLE WITH RFC[1000]