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Team:KULeuven/Sandbox
From 2009.igem.org
Sandbox
Team:KULeuven/Sandbox/Design1 Team:KULeuven/Sandbox/Design2
Blue Light Receptor
The receptor senses the blue light to which the bacteria are exposed. By choosing the light intensity, the wanted vanillin concentration is set. Upon photo-excitation the receptorprotein YcgF dimerizes and interacts directly with the repressor YcgE. This protein is bound to the promoter-region of the key-gene and inhibits its transcription. The dimerized YcgF acts as an anti-repressor and releases YcgE from the DNA. Thereupon, the transcribed key activates vanillin production, which is proportional to the entered intensity of the blue light.
Vanillin Receptor
The vanillin receptor senses the vanillin concentration outside the cell. This information is needed to create the feedback loop that controls vanillin synthesis. The receptor consists of two proteins: virA and virG. In the presence of vanillin, virA binds a phosphor and transfers it to virG. In turn, the phosphorylated VirG binds to a so-called vir box sequence, triggering transcription of antikey. The more antikey is produced, the more it can anneal to the key and the less vanillin is produced. This process goes on until there’s an equilibrium between measured and wanted concentration.
Key Lock Antikey
The key/antikey system compares the signal from the blue light and vanillin receptor in order to control vanillin production. The more the measured amount of vanillin exceeds the wanted amount (set by the blue light intensity), the less vanillin is produced. After blue light irradiation, the key is transcribed and ‘unlocks’ the vanillin synthesis pathway. In response to the produced vanillin, the vanillin receptor then activates transcription of the antikey. Key and antikey are complementary RNA-strands: their annealing is favoured over the reaction between key en lock leading to vanillin synthesis.
Vanillin Production
A vanilla odour is created by synthesizing the molecule Vanillin. The starting point is tyrosine, an amino acid produced endogenously in E.coli. The subsequent pathway involves a combination of five enzymes. By locking both the transcription of the first and the third enzyme we prevent vanillin synthesis without the presence of the key.
