Part:BBa_K2042001

Pseudomonas resinovorans PHA synthase 1

This is the coding sequence for a class II poly(R)-hydroxyalkanoic acid (PHA) synthase 1 from Pseudomonas resinovorans. The enzyme takes the monomer (D)-lactyl-CoA and produce the PLA homopolymer. This sequence is codon optimized for Pseudomonas putida and includes four amino acid mutations (E130D/S325T/S477G/Q481K) which were found to increase yields of PLA. This type II PHA synthase 1 accept short-chain-length hydroxyacyl-CoAs including lactyl-CoA and 3-hydroxybutyryl-CoA as substrates and support the synthesis of P(3HB-co-LA). This biobrick takes over Yale 2013 BBa_K1211002 biobrick. Indeed according to "http://www.ncbi.nlm.nih.gov/pubmed/21221571" the mutation on serine 477 is needed for PLA polymerization but in BBa_K1211002 this mutation is found on serine 475.

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 87
  Illegal XhoI site found at 172
  Illegal XhoI site found at 1030
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 1060
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 796