Team:KULeuven/Lab/Vanillin Production

=VANILLIN SYNTHESIS=

Goal
Making vanillin from tyrosine in a five-step pathway.

Required
Biobricks:
 * SAM8: : coding sequence without RBS
 * SAM5: : RBS site + PstI restriction site removed
 * COMT: : coding sequence without RBS
 * FCS: : RBS + fcs in pSB1A2
 * ECH: : RBS + ech in pSB1A2

Where from

 * 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 ferrulic acid. The enzymes are put under a constitutive promoter and transcribed. Ferrulic acid concentrations are then measured with GC
 * 2) Testing the transformation of ferrulic acid to vanillin. The enzymes are put under a constitutive promoter and transcribed. Ferrulic 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 tyrosine of E. coli is depleted too fast