Difference between revisions of "Part:BBa K2042000"

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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.  
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This is an evolved coding sequence for the enzyme Propionate CoA transferase (Pct) EC 2.8.3.1. The wild type enzyme comes 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 [1,2]. Moreover, this sequence is codon optimized for Pseudomonas putida using Codon Optimization Tool from IDT in order to produce Polylactic acid (PLA).  
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This enzyme is a variant of Yale 2013 <html><a href="/Part:BBa_K1211001">BBa_K1211001</a></html> 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.
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<b>This part can be considered as an improvement of the biobrick Clostridum propionicum propionate CoA transferase (BBa_K1211001)</b>: BBa_K1211001 is designed to work in Pseudomonas putida, one different amino acid substitution (A243T instead of V193A) where used, which also led to the increase of both polymer content and lactate mole fraction in the copolymer.
  
 
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<partinfo>BBa_K2042000 parameters</partinfo>
 
<partinfo>BBa_K2042000 parameters</partinfo>
 
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===Predicted normalized B-factor===
 
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[[File:BBa K2042000 BFP.png|caption]]
 
  
 
===Predicted structure===
 
===Predicted structure===
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[[File:BBa_K2042000_model.gif|frameless|Predicted structure of pct by I-TASSER]]
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[[File:BBa_K2042000_model.gif|none|upright=0.9|left|Predicted structure of pct by I-TASSER]]
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===References===
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[1] Biosynthesis of Polylactic Acid and Its Copolymers using Evolved Propionate CoA Transferase and PHA Synthase <br>
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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 <br>
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http://www.ncbi.nlm.nih.gov/pubmed/19937726
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[2] Microbial production of lactate-containing polyesters. <br>
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Yang JE1, Choi SY, Shin JH, Park SJ, Lee SY.<br>
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http://www.ncbi.nlm.nih.gov/pubmed/23718266

Latest revision as of 00:33, 30 October 2016


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 comes 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 [1,2]. Moreover, this sequence is codon optimized for Pseudomonas putida using Codon Optimization Tool from IDT in order to produce Polylactic acid (PLA). This part can be considered as an improvement of the biobrick Clostridum propionicum propionate CoA transferase (BBa_K1211001): BBa_K1211001 is designed to work in Pseudomonas putida, one different amino acid substitution (A243T instead of V193A) where used, which also led to the increase of both polymer content and lactate mole fraction in the copolymer.

Part_BBa_K2042000.jpeg

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

[1] 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

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