Additional Ideas for Future Programmed Evolution Research

1. Develop a multistep pathway off of the caffeine->theophylline single step pathway (Find a substrate that creates caffeine)

2. Thiamine pyrophosphate riboswitch File:Thiamine Pyrophosphate Riboswitch.pdf

  -In E.coli, thiamine monophosphate is converted to thiamine which then could be converted to thiamine pyrophosphate by thiamine kinase. For this particular riboswitch when thiamine pyrophosphate (TPP) binds to the riboswitch, it will stop transcription. A discussion about the thiamine pyrophosphate riboswitch resulted in the decision to not pursue this specific riboswitch because E.coli relies on this metabolic system to survive and it could be impossible to remove thiamine and still have them survive. This riboswitch is a great example of a multi-step metabolic pathway, but unfortunately it would be too detrimental to remove thiamine from E.coli’s system. It is still a goal to research a multi-step pathway as fascinating as the thiamine->>thiamine pyrophosphate, but still remain similar and practical to the caffeine->theophylline system.