Part:BBa_K3590042

Enzyme MlrC derived from Sphyngopyxis sp. fused with a dockerin derived from Acetivibrio cellulolyti

MlrC is the third enzyme of the microcystin degradation pathway. The purpose of MlrC-DocAc is to further degrade the products of MlrA and MlrB enzymatic activity into even shorter linear peptides and amino acids (one of its products is ADDA). It thus contributes to the reduction of microcystin induced toxicity. The enzyme should fulfill its task while being displayed on the ScafD.

At the 5' end of MlrC-DocAc there the Pveg promoter followed by the RBS R0. The CDS encodes several functional modules - SacB signal sequence for extracellular transport, His-tag for Western blot detection followed by the MlrC module, the function of which is described above. MlrC is connected by a short linker to the dockerin from Acetivibrio cellulolyticus. This dockerin interacts exclusively with a cohesin domain originating from the same species so it ensures the attachment of this fusion protein to the specific position on ScafD. At the 3' end of the CDS, there is a STOP codon.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 596
  Illegal BglII site found at 753
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 124
  
• 1000
  COMPATIBLE WITH RFC[1000]