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Team:KULeuven/Lab/Vanillin Production
From 2009.igem.org
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==required== | ==required== | ||
biobricks: | biobricks: | ||
| - | *SAM8: | + | *SAM8: {{kulpart|BBa_I742142}}: coding sequence without RBS |
| - | *SAM5: BBa_K238007: RBS site + PstI restriction site removed | + | *SAM5: {{kulpart|BBa_K238007}}: RBS site + PstI restriction site removed |
| - | *COMT: BBa_I742107: coding sequence without RBS | + | *COMT: {{kulpart|BBa_I742107}}: coding sequence without RBS |
| - | *FCS: BBa_I742115: RBS + fcs in pSB1A2 | + | *FCS: {{kulpart|BBa_I742115}}: RBS + fcs in pSB1A2 |
| - | *ECH: BBa_I742113: RBS + ech in pSB1A2 | + | *ECH: {{kulpart|BBa_I742113}}: RBS + ech in pSB1A2 |
==Where from== | ==Where from== | ||
Revision as of 15:43, 28 July 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
