Difference between revisions of "Part:BBa K1149002"
Line 6: Line 6:
 
The enzyme is the estCS2 from [https://parts.igem.org/Part:BBa_K892012 BBa_K892012] previous biobrick by Washington 2012 team, that encodes carboxylesterase that breaks down ester bonds in polyurethane. We have added a [https://parts.igem.org/Part:BBa_K1149023 pelB] seretion tag to the enzyme so that it can be secreted extacellularly. We also added a 6xHistidine tag for Western blotting and protein purification purposes.  
 
The enzyme is the estCS2 from [https://parts.igem.org/Part:BBa_K892012 BBa_K892012] previous biobrick by Washington 2012 team, that encodes carboxylesterase that breaks down ester bonds in polyurethane. We have added a [https://parts.igem.org/Part:BBa_K1149023 pelB] seretion tag to the enzyme so that it can be secreted extacellularly. We also added a 6xHistidine tag for Western blotting and protein purification purposes.  
  
−
Theoretical pathway of PUR degradation by esterase enzymes:
+
Theoretical pathway of PUR degradation by esterase:
 
https://static.igem.org/mediawiki/igem.org/f/fd/PUR_esterase_theoretical_pathway.jpg
 
https://static.igem.org/mediawiki/igem.org/f/fd/PUR_esterase_theoretical_pathway.jpg
  
  
−
references:  
+
<p>references: </p>
−
[http://www.selu.edu/acad_research/depts/biol/faculty/publications/pdf/2012/howard2012.pdf Howard, Gary yr:2012 pg:371 -3, Polyurethane Biodegradation, Environmental Science [1431-6250]]
+
[http://www.selu.edu/acad_research/depts/biol/faculty/publications/pdf/2012/howard2012.pdf Howard, Gary 2012 pg:371 -3, Polyurethane Biodegradation, Environmental Science]
  
−
[http://www.microbialcellfactories.com/content/10/1/41 A novel family VII esterase with industrial potential from compost metagenomic library. Microbial Cell Factories 2011, 10:41 doi:10.1186/1475-2859-10-41]
+
[http://www.microbialcellfactories.com/content/10/1/41 A novel family VII esterase with industrial potential from compost metagenomic library. Microbial Cell Factories 2011]
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 17:24, 15 September 2013

pelB-EstCS2-his expression (GFP reporter)

This part expresses a plastic degradation enzyme and GFP under the control of an arabinose inducible promoter.

The enzyme is the estCS2 from BBa_K892012 previous biobrick by Washington 2012 team, that encodes carboxylesterase that breaks down ester bonds in polyurethane. We have added a pelB seretion tag to the enzyme so that it can be secreted extacellularly. We also added a 6xHistidine tag for Western blotting and protein purification purposes.

Theoretical pathway of PUR degradation by esterase: PUR_esterase_theoretical_pathway.jpg


references:

[http://www.selu.edu/acad_research/depts/biol/faculty/publications/pdf/2012/howard2012.pdf Howard, Gary 2012 pg:371 -3, Polyurethane Biodegradation, Environmental Science]

[http://www.microbialcellfactories.com/content/10/1/41 A novel family VII esterase with industrial potential from compost metagenomic library. Microbial Cell Factories 2011]

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
    Illegal BamHI site found at 692
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 210
    Illegal NgoMIV site found at 958
    Illegal NgoMIV site found at 1192
    Illegal AgeI site found at 2131
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1006