Coding

Part:BBa_K4886000:Design

Designed by: Ruyi Shi   Group: iGEM23_Nanjing-BioX   (2023-07-03)


Phosphoketolase(FXpk) from Bifidobacterium adolescentis


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 1675
    Illegal PstI site found at 299
    Illegal PstI site found at 1970
    Illegal PstI site found at 2450
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 1675
    Illegal PstI site found at 299
    Illegal PstI site found at 1970
    Illegal PstI site found at 2450
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 1675
    Illegal PstI site found at 299
    Illegal PstI site found at 1970
    Illegal PstI site found at 2450
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 1675
    Illegal PstI site found at 299
    Illegal PstI site found at 1970
    Illegal PstI site found at 2450
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

It is a key enzyme in the NOG pathway. 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.

Figure 1 NOG pathway and related enzyme genes (red arrows indicate lack of key enzymes in Clostridium tyrobutyricum)

Source

The gene is derived from Clostridium acetobutylicum ATCC824.

References