Part:BBa_K4885010
10
This part is responsible for the expression of adhE2, NGbcd (BBa_K4885007) and NGcrt (BBa_K4885008) with Pthl promoter. adhE2 gene is derived from Clostridium acetobutylicum and codes for the alcohol/aldehyde bifunctional dehydrogenase. NGbcd contains NGCatcher sequence and bcd gene and codes for fusion protein of NGCatcher and butyryl-CoA dehydrogenase. NGcrt contains NGTag sequence and crt gene and codes for NGTag-crotonase fusion protein. Bcd and crt genes are derived from Clostridium tyrobutyricum (C. tyrobutyricum) L319. Alcohol/aldehyde bifunctional dehydrogenase converts acyl-CoA to aldehyde and then to alcohol in two reductive steps. Crotonase converts 3-hydroxybutyryl-CoA to crotonyl-CoA, and butyryl-CoA dehydrogenase converts crotonyl-CoA to butyryl-CoA in the butyrate synthesis pathway in C. tyrobutyricum. NGTag forms a covalent bond with NGCatcher rapidly which is not affected by the change of environment. Butyryl-CoA dehydrogenase (bcd) and crotonase (crt) modified by NGCatcher/NGTag oligopeptides can be assembled together. Pthl is a strong transcriptional promoter in C. tyrobutyricum.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Results
(1)Plasmid construction and transformation
(2)Product yields of the engineered C. tyrobutyricum
None |