Part:BBa_K1060003

Methyl Salicylate Production

KU_Leuven 2013 requested the BBa_J45700 brick from MIT 2006 and recloned into the pSB1C3 standard plasmid backbone. For the assembly of this part we refer to the original MIT brick BBa_J45700 => "We assembled a complete wintergreen odor biosynthetic system (BBa_J45700) by combining the salicylic acid (BBa_J45320) and wintergreen odor generators (BBa_J45120). BBa_J45700 catalyzes the conversion of the cellular metabolite chorismate to methyl salicylate or wintergreen odor."

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 744
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 1689
  Illegal NgoMIV site found at 2093
  Illegal NgoMIV site found at 2218
  Illegal NgoMIV site found at 2229
  Illegal NgoMIV site found at 2510
  Illegal NgoMIV site found at 2978
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 285
  

Headspace Gas-Chromatography

Samples were analyzed with a calibrated Autosystem XL gas chromatograph with a headspace sampler (HS40; Perkin-Elmer, Wellesley, Mass.) and equipped with a CP-Wax 52 CB column. Headspace GC was performed on pure MeS (Sigma-Aldrich) (A), on culture supernatant from a E. coli BL21 strain harbouring BBa_K1060003 without (B) or with 0.1 mM salicylic acid (C). MeS could be detected at 12.832 minutes in (A) and (C) but not when no salicylic acid was added. (For more details please go to [http://2013.igem.org/Team:KU_Leuven/Project/Glucosemodel/MeS here].)