Composite

Part:BBa_K4885012:Design

Designed by: Leyu Xu   Group: iGEM23_Nanjing-SDG   (2023-10-09)


Ptkt-adhE2


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We designed Ptkt-adhE2 to improve the part Pthl-adhE2. Clostridium tyrobutyricum (C. tyrobutyricum) transfected with the plasmid containing Pthl-adhE2 had the capacity to produce butanol in addition to its native capacity of butyrate synthesis, which can be used to manufacture butyl butyrate by lipase catalyzed esterification. For the esterification reaction, the optimal butyrate-to butanol molar ratio is 1:1, however this strain produced far more butanol than butyrate. Here, we used a weaker promoter Ptkt to express adhE2 to lower the yield of butanol and increase the yield of butyrate in the strain, so the engineered strain could achieve a better production ratio of the two precursors for butyl butyrate synthesis.

Source

Ptkt sequence is from BBa_K4886005; adhE2 sequence is from BBa_K1462060; terminator sequence is from BBa_K3585002

References