Coding

Part:BBa_K2042001

Designed by: Toky RATOVOMANANA   Group: iGEM16_Evry   (2016-09-02)
Revision as of 15:32, 4 September 2016 by RATOVOMANANA (Talk | contribs)


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


[edit]
Categories
//cds/biosynthesis
//chassis/prokaryote/pputida
Parameters
None