Part:BBa_K4886001:Design
Pthl-F/Xpk(BD)
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1986
Illegal XbaI site found at 929 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1986
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1986
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1986
Illegal XbaI site found at 929 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1986
Illegal XbaI site found at 929 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
Our project aims to reduce the carbon emission during the fermentation of Clostridium tyrobutyricum (C. tyrobutyricum) by constructing a NOG pathway to integrate with the native EMP pathway of the strain. The NOG pathway is an artificial pathway and functions as a fructose-6-phosphate (F6P) shunt. Through carbon rearrangement, this pathway can convert 1mol of F6P to 3mol of AcP without any loss of carbon. By comparison, we found that most of the key enzymes in the NOG pathway natively exist in C. tyrobutyricum and the functions of the absent key enzymes can be carried out by phosphoketolases (Figure 1). Therefore, in order to construct the NOG pathway in C. tyrobutyricum L319, we introduced phosphoketolase (F/Xpk) gene into the strain. Pthl promoter is a strong transcriptional promoter in C. tyrobutyricum L319. We used this promoter to ensure a robust expression in the strain.
