Difference between revisions of "Part:BBa K4886006:Design"

 
(→‎Design Notes)
Line 7: Line 7:
  
 
===Design Notes===
 
===Design Notes===
−
1
+
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. Ptkt promoter is a native transcriptional promoter in C. tyrobutyricum L319. We used this promoter, Pthl and Pfba promoters to test which can provide the most  robust expression of F/Xpk in the strain.
−
 
+
Figure 1 NOG pathway and related enzyme genes (red arrows indicate lack of key enzymes in Clostridium tyrobutyricum)
−
 
+
  
 
===Source===
 
===Source===

Revision as of 09:30, 25 September 2023


Ptkt-P/Xpk(BD)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1675
    Illegal XbaI site found at 618
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1675
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1675
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1675
    Illegal XbaI site found at 618
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1675
    Illegal XbaI site found at 618
  • 1000
    COMPATIBLE 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. Ptkt promoter is a native transcriptional promoter in C. tyrobutyricum L319. We used this promoter, Pthl and Pfba promoters to test which can provide the most robust expression of F/Xpk in the strain. Figure 1 NOG pathway and related enzyme genes (red arrows indicate lack of key enzymes in Clostridium tyrobutyricum)

Source

1

References