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

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===Design Notes===
 
===Design Notes===
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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.
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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===
 
===Source===

Revision as of 06:47, 30 September 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_K4119027

crt sequence is from BBa_K4119028

Cpa fdx terminator sequence is from BBa_K3585002

References