Difference between revisions of "Part:BBa K4885005:Design"

Latest revision as of 13:05, 12 October 2023

Pthl-bcd-crt

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 798
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE 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

@@ Line 14: / Line 14: @@
 <p>Pthl sequence is from BBa_K3443002</p>
 <p>RBS for adhE2 sequence is from BBa_K4408001</p>
-<p>bcd sequence is from BBa_K4119027</p>
+<p>bcd sequence is from BBa_K4885001</p>
 <p>crt sequence is from BBa_K4119028</p>
 <p>Cpa fdx terminator sequence is from BBa_K3585002</p>
 ===References===