Composite

Part:BBa_K1149005

Designed by: Margarita Kopniczky   Group: iGEM13_Imperial_College   (2013-09-14)
Revision as of 19:10, 15 September 2013 by Margarita K (Talk | contribs)

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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]


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Parameters
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