Part:BBa_K2042000

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 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 Polylactic acid (PLA) in P. putida. This enzyme is a variant of Yale 2013 BBa_K1211001 biobrick but codon optimized for Pseudomonas putida and with one different amino acid mutation (A243T instead of V193A) which also led to the increase of both polymer content and lactate mole fraction in the copolymer.

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
  

Predicted structure

Predicted structure of pct by I-TASSER

References

Biosynthesis of Polylactic Acid and Its Copolymers using Evolved Propionate CoA Transferase and PHA Synthase
Taek Ho Yang,1 Tae Wan Kim,1 Hye Ok Kang,1 Sang-Hyun Lee,1 Eun Jeong Lee,1 Sung-Chul Lim,1 Sun Ok Oh,1 Ae-Jin Song,2 Si Jae Park,1 Sang Yup Lee2,3
http://www.ncbi.nlm.nih.gov/pubmed/19937726

Microbial production of lactate-containing polyesters.
Yang JE1, Choi SY, Shin JH, Park SJ, Lee SY.
http://www.ncbi.nlm.nih.gov/pubmed/23718266