Difference between revisions of "Part:BBa K2042001:Design"

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===Design Notes===
 
===Design Notes===
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We used this sequence as second enzyme in order to produce
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This is the DNA coding for the enzyme Polyhydroxyalkanoate synthase from Pseudomonas resinovorans. This sequence was codon optimized for Pseudomonas putida and four amino acid mutations were introduced to increase yields of Polylactic acid (PLA). Silent mutation were also introduced for removing restriction site.
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PLA. In fact it will take lactyl coA as substract and polymerise it into PLA homopolymere
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The iGEM Team Evry 2016 used this enzyme as the second enzyme in the pathway to produce PLA in Pseudomonas putida.
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===Source===
 
===Source===
  
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Pseudomonas resinovorans gDNA.
 
This sequence was synthetized by IDT
 
This sequence was synthetized by IDT
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===References===
 
===References===
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Biosynthesis of Polylactic Acid and Its Copolymers using Evolved Propionate CoA Transferase and PHA Synthase
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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
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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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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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Tailor-made type II Pseudomonas PHA synthases and their use for the biosynthesis of polylactic acid and its copolymer in recombinant Escherichia coli.
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Microbial production of lactate-containing polyesters. <br>
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Yang TH1, Jung YK, Kang HO, Kim TW, Park SJ, Lee SY.
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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 17:39, 8 October 2016


Pseudomonas resinovorans PHA synthase 1


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


Design Notes

This is the DNA coding for the enzyme Polyhydroxyalkanoate synthase from Pseudomonas resinovorans. This sequence was codon optimized for Pseudomonas putida and four amino acid mutations were introduced to increase yields of Polylactic acid (PLA). Silent mutation were also introduced for removing restriction site. The iGEM Team Evry 2016 used this enzyme as the second enzyme in the pathway to produce PLA in Pseudomonas putida.

Source

Pseudomonas resinovorans gDNA. This sequence was synthetized by IDT

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

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

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