Difference between revisions of "Part:BBa K1149005"
 
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<partinfo>BBa_K1149005 short</partinfo>
 
<partinfo>BBa_K1149005 short</partinfo>
  
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This part can express a plastic degradation enzyme from Comamonas acidovorans that breaks down ester bonds in polyurethane. The sequence was codon optimized for E.coli. We added a pelB tag to ensure secretion to the N terminal and we have added a 6xpolyhistidine tag to the C terminal.  
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This part can express a plastic degradation enzyme from Comamonas acidovorans that breaks down ester bonds in polyurethane. It containes a Gly-X-Ser-X-Gly motif characteristic of serine hydrolases. The sequence was codon optimized for E.coli. We added a pelB tag to ensure secretion to the N terminal and we have added a 6xpolyhistidine tag to the C terminal.  
 
The promoter is arabinose inducible.  
 
The promoter is arabinose inducible.  
  
  
 
Theoretical pathway of PUR degradation by esterase:
 
Theoretical pathway of PUR degradation by esterase:
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https://static.igem.org/mediawiki/igem.org/f/fd/PUR_esterase_theoretical_pathway.jpg
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https://static.igem.org/mediawiki/parts/3/31/PUR_esterase_theoretical_pathway_2.jpg
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<p>references: </p>
 
<p>references: </p>
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[http://www.selu.edu/acad_research/depts/biol/faculty/publications/pdf/2012/howard2012.pdf Howard, Gary 2012 pg:371 -3, Polyurethane Biodegradation, Environmental Science]
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<p> [http://www.sciencedirect.com/science/article/pii/S0922338X99890011 Cloning and sequence analysis of a polyurethane esterase of Comamonas acidovorans 1998] </p>
 
<p> [http://www.sciencedirect.com/science/article/pii/S0922338X99890011 Cloning and sequence analysis of a polyurethane esterase of Comamonas acidovorans 1998] </p>
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<p> [http://www.sciencedirect.com/science/article/pii/S0964830598000663Purification and characterization of a soluble polyurethane degrading enzyme from Comamonas acidovorans 1999 ] </p>
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<p> [http://www.sciencedirect.com/science/article/pii/S0964830598000663 Purification and characterization of a soluble polyurethane degrading enzyme from Comamonas acidovorans 1999] </p>
  
  

Latest revision as of 19:10, 15 September 2013

pelB-pudA expression

This part can express a plastic degradation enzyme from Comamonas acidovorans that breaks down ester bonds in polyurethane. It containes a Gly-X-Ser-X-Gly motif characteristic of serine hydrolases. The sequence was codon optimized for E.coli. We added a pelB tag to ensure secretion to the N terminal and we have added a 6xpolyhistidine tag to the C terminal. The promoter is arabinose inducible.


Theoretical pathway of PUR degradation by esterase: PUR_esterase_theoretical_pathway_2.jpg

references:

[http://www.sciencedirect.com/science/article/pii/S0922338X99890011 Cloning and sequence analysis of a polyurethane esterase of Comamonas acidovorans 1998]

[http://www.sciencedirect.com/science/article/pii/S0964830598000663 Purification and characterization of a soluble polyurethane degrading enzyme from Comamonas acidovorans 1999]


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 125
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 65
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 529
    Illegal AgeI site found at 604
    Illegal AgeI site found at 1753
  • 1000
    COMPATIBLE WITH RFC[1000]