Part:BBa_K1983000

Codon optimized AvPAL with C-terminal 6XHis-Tag

Phenylalanine ammonia lyase (PAL) from Anabaena variabilis is an enzyme that catalyzes breaking down L-phenylalanine to ammonia and trans-cinnamic acid. This is an improved Stanford-Brown 2013 AvPAL biobrick part. We have removed the T7 promoter and replaced the N-terminal FLAG-tag with a more common C-terminal 6xHis-tag. The C-terminal tag location was chosen assuming it would less likely affect the stability and folding of the protein.

Our team improved the characterization of this part by testing the enzyme's activity in vivo. We also have experimentally measured the expression rate and time to reach steady of the protein in Escherichia coli cells, since the kinetic constants of this enzyme are determined[]. Expression of the 6XHis-tagged protein is easily detectable via Western Blot. We used pBAD expression system for our project and characterization of this part.

Experiments and Results

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 1265
  Illegal XhoI site found at 1702
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1483
  
• 1000
  COMPATIBLE WITH RFC[1000]