Difference between revisions of "Part:BBa K4885001:Design"

 
(→‎Design Notes)
 
(One intermediate revision by the same user not shown)
Line 7: Line 7:
  
 
===Design Notes===
 
===Design Notes===
−
1
+
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 are considered as rate-limiting enzymes.
−
 
+
We used the endogenous bcd and crt genes and Pthl promoter from C. tyrobutyricum L319 to construct the part of Pthl-bcd-crt to overexpress crt and bcd in C. tyrobutyricum to enhance the butyrate and butanol synthesis pathway.
−
 
+
  
 
===Source===
 
===Source===
  
−
1
+
The part sequence is obtained by mutating two sites from BBa_K4119027
  
 
===References===
 
===References===

Latest revision as of 13:14, 1 October 2023


bcd,butyryl-CoA dehydrogenase coding sequence


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 181
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 310


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 are considered as rate-limiting enzymes. We used the endogenous bcd and crt genes and Pthl promoter from C. tyrobutyricum L319 to construct the part of Pthl-bcd-crt to overexpress crt and bcd in C. tyrobutyricum to enhance the butyrate and butanol synthesis pathway.

Source

The part sequence is obtained by mutating two sites from BBa_K4119027

References