Part:BBa_K4885005:Design
Pthl-bcd-crt
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 798
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1977
Illegal SapI.rc site found at 927
Design Notes
In C. tyrobutyricum L319, glucose is converted to acetyl-CoA which is then mainly converted to butyryl-CoA by the following enzymes: thiolase (thl), 3-hydroxybutyryl-CoA dehydrogenase (hbd), crotonase (crt), and butyryl-CoA dehydrogenase (bcd). Butyryl-CoA is subsequently converted to butyrate in the native strain. In the preliminary experiment, we discovered that the yield of butyrate and butanol increased efficiently through the overexpression of bcd and crt. Therefore, crotonase and butyryl-CoA dehydrogenase coded by bcd and crt are considered as rate-limiting enzymes. Here, we used the endogenous bcd and crt genes and Pthl promoter from C. tyrobutyricum L319 to construct the part of Pthl-bcd-crt and the recombinant plasmid of pMTL-adhE2-bcd-crt to overexpress crt and bcd in C. tyrobutyricum to enhance the butyrate and butanol synthesis pathway.
Source
Pthl sequence is from BBa_K3443002
RBS for adhE2 sequence is from BBa_K4408001
bcd sequence is from BBa_K4885001
crt sequence is from BBa_K4119028
Cpa fdx terminator sequence is from BBa_K3585002