Team:KULeuven/Lab/Vanillin Production

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VANILLIN SYNTHESIS

Goal

Making vanillin from tyrosine in a five-step pathway.

Making vanillin from tyrosine

required

  • biobricks: 
   SAM8: BBa_ I742142: coding sequence without RBS
   SAM5: no number: RBS site + PstI restriction site removed
   COMT: BBa_I742107: coding sequence without RBS
   FCS: BBa_I742115: RBS + fcs in pSB1A2 
   ECH: BBa_I742113: RBS + ech in pSB1A2

Where from

  • 1. COMT is available in 2009 spring distribution plate 1 well 19D

DNA is compatible

  • 2. SAM8, SAM5, COMT, FCS, ECH will be sent to us from the French Lab from the university Edinburgh

Steps

  • we will be working in three different stages
  1. Testing the transformation of tyrosin to ferulic acid. The enzymes are put under a constitutive promoter and transcribed. Ferulic acid concentrations are then measured with GC
  2. Testing the transformation of ferulic acid to vanillin. The enzymes are put under a constitutive promoter and transcribed. Ferulic acid concentrations are then measured with GC
  3. If both are tested and ok, they can be combined into one plasmid and again tested.
  4. Then, the locks need to be added.
  • testing the enzymes: by detecting the endproducts through gaschromatography.
  • One problem might be that the tyrosin of E.Coli might be depleted to fast
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