Composite
Part:BBa_K4885008:Design
Designed by: Leyu Xu Group: iGEM23_Nanjing-SDG (2023-09-10)
8
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 214
Design Notes
In the butyrate synthesis pathway of C. tyrobutyricum L319, crotonase (crt) converts 3-hydroxybutyryl-CoA to crotonyl-CoA which is subsequently converted to butyryl-CoA by butyryl-CoA dehydrogenase coded by bcd. These two enzymes are rate-limiting enzymes. We used this part and BBa_K4885007 to express fusion protein of NGTag and crotonase (crt) and fusion protein of NGCatcher and butyryl-CoA dehydrogenase (bcd) in C. tyrobutyricum, which allows the assembly of crotonase and butyryl-CoA dehydrogenase via the NGTag/NGCatcher peptide interaction. In this way, we were able to enhance the butyrate synthesis pathway in the strain.
Source
NGTag sequence is from BBa_K4119030 crt sequence is from BBa_K4119028.