Design
There are several metabolic pathways for the biosynthesis of vanillin.
1）Ferulic acid → Feruloyl-CoA or 4-Vinylguaiacol → Vanillin
2）Phenylalanine → Ferulic acid → Vanillin
3）Glucose → 3-dehydrosikimic acid → Vanillin
We chose this metabolic pathway starting from glucose. From glucose, we metabolize it to L-phenylalanine with 3-dehydrosikimic acid, and from L-phenylalanine we incorporate an enzyme gene to metabolize it to vanillin.
The vector is bba_j04450 and the following enzyme genes are incorporated into it.
BBa_j04450 has a Lac promoter, which allows it to control vanillin production. （Figure）
We use genetic modification to biosynthesize vanillin, but to determine if vanillin is actually being produced, we use electrophoresis to confirm that the genes are correctly incorporated and detect whether the compounds leading up to vanillin are being synthesized in the gas mass, or whether the end product vanillin is being synthesized.
