Part:BBa_K2042001

Pseudomonas resinovorans PHA synthase 1

This is the coding sequence for a Polyhydroxyalkanoate (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 it includes four mutations which were found to increase yields of PLA. The type II PHA synthase 1 is engineered to accept short-chain-length hydroxyacyl-CoAs including lactyl-CoA and 3-hydroxybutyryl-CoA as substrates and support the synthesis of P(3HB-co-LA) by site-directed mutagenesis of four sites, E130, S325, S477, and Q481 One of the mutation substitue the serine 477 in G differently in We note that unlike BioBrick BBa K1211002 Yale 2013, one of the mutation is on the serine 477 not on serine 475 which is wrong. The enzyme is able to do the reaction: lactylCoA + PLA(n) = PLA(n+1).

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