Difference between revisions of "Part:BBa K2042001"
 
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<partinfo>BBa_K2042001 short</partinfo>
 
<partinfo>BBa_K2042001 short</partinfo>
  
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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.
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This sequence is codon optimized for pseudomonas putida it includes four mutations which were found to increase yields of PLA.
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This is the coding sequence for 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.
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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
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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 [1,2].
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One of the mutation substitue the serine 477 in G differently in
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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)[3].
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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.
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This biobrick correct Yale 2013 <html><a href="https://parts.igem.org/Part:BBa_K1211002" title="BBa_K1211002">BBa_K1211002</a></html> biobrick. Indeed according to "http://www.ncbi.nlm.nih.gov/pubmed/21221571"  the mutation on serine 477 is needed for PLA polymerization but in <html><a href="https://parts.igem.org/Part:BBa_K1211002" title="BBa_K1211002">BBa_K1211002</a></html> this mutation is found on serine 475.
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The enzyme is able to do the reaction: lactylCoA + PLA(n) = PLA(n+1).
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<!-- Add more about the biology of this part here
 
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===Usage and Biology===
 
===Usage and Biology===
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The enzyme is able to do the reaction: lactylCoA + PLA(n) = PLA(n+1).
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<span class='h3bb'>Sequence and Features</span>
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<span class='h3bb'><h3>Sequence and Features</h3></span>
 
<partinfo>BBa_K2042001 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2042001 SequenceAndFeatures</partinfo>
  
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<partinfo>BBa_K2042001 parameters</partinfo>
 
<partinfo>BBa_K2042001 parameters</partinfo>
 
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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] Tailor-made type II Pseudomonas PHA synthases and their use for the biosynthesis of polylactic acid and its copolymer in recombinant Escherichia coli. <br>
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Yang TH1, Jung YK, Kang HO, Kim TW, Park SJ, Lee SY.<br>
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http://www.ncbi.nlm.nih.gov/pubmed/21221571
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[3] 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 00:28, 30 October 2016


Pseudomonas resinovorans PHA synthase 1


This is the coding sequence for 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 [1,2]. 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)[3]. This biobrick correct 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


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] Tailor-made type II Pseudomonas PHA synthases and their use for the biosynthesis of polylactic acid and its copolymer in recombinant Escherichia coli.
Yang TH1, Jung YK, Kang HO, Kim TW, Park SJ, Lee SY.
http://www.ncbi.nlm.nih.gov/pubmed/21221571

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