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Team:KULeuven/Lab/Vanillin Production
From 2009.igem.org
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[[Image:Vanillin_Biosynthesis_Pathway.jpg|center|thumb|365px|Making vanillin from tyrosine]] | [[Image:Vanillin_Biosynthesis_Pathway.jpg|center|thumb|365px|Making vanillin from tyrosine]] | ||
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*SAM8: {{kulpart|BBa_I742142}}: coding sequence without RBS | *SAM8: {{kulpart|BBa_I742142}}: coding sequence without RBS | ||
*SAM5: {{kulpart|BBa_K238007}}: RBS site + PstI restriction site removed | *SAM5: {{kulpart|BBa_K238007}}: RBS site + PstI restriction site removed | ||
Revision as of 11:21, 10 August 2009
Contents |
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
- 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
- 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
- If both are tested and ok, they can be combined into one plasmid and again tested.
- 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
