Coding
Pct

Part:BBa_K2042000

Designed by: Toky RATOVOMANANA   Group: iGEM16_Evry   (2016-08-19)
Revision as of 19:22, 29 August 2016 by RATOVOMANANA (Talk | contribs)


Propionate CoA transferase for P. putida

This is an evolved coding sequence for the enzyme Propionate CoA transferase (Pct) EC 2.8.3.1 . The wild type enzyme come from Clostridium propionicum and normally catalyzes the reaction: acetyl-CoA + propanoate <--> acetate + propanoyl- CoA. This mutated sequence has one amino acid substitution: A243T allowing lactate to act as the acceptor and convert it efficiently to lactyl-CoA. Moreover, this sequence is codon optimized for Pseudomonas putida using Codon Optimization Tool from IDT in order to produce (D)-Lactyl- CoA. This enzyme is a variant of Yale 2013 BBa_K1211001 biobrick codon optimized for Pseudomonas putida and with one different amino acid mutation (A243T instead of V193A) which led to the increase of both polymer content and lactate mole fraction in the copolymer.


Figure 1 Reaction of the .

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 675
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1081
    Illegal BsaI site found at 1552


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