Difference between revisions of "Part:BBa K5236013"

 
 
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<partinfo>BBa_K5236013 short</partinfo>
 
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This basic part encodes for a BhrPETase LCCICCG and is derived from Escherichia coli.  
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Cutinase (EC 3.1.1.74) is a lipolytic/esterolytic enzyme that hydrolyzes not only cutin, which is a major component of plant cuticl, but also water-soluble esters and insoluble triglycerides. Leaf-branch compost cutinase (LCC) was first reported in 2012 and its activity and thermostability improved via four mutations yielding LCC-ICCG.  
  
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===Usage and Biology===
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We tested for successful plasmid construction and transformation into E.coli through colony PCR and gel electrophoresis. The following gel result demonstrates that the plasmid transformed into E.coli are correct. The plasmid should have a total of 891 base pairs and the results match.
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<center><html><img src ="https://static.igem.wiki/teams/5236/part-images/colony-pcr.png" width = "50%"><br></html></center>
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<center>Fig.1 The DNA gel electrophoresis result </center>
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<center><html><img src ="https://static.igem.wiki/teams/5236/part-images/lcciccg-sequence.png" width = "50%"><br></html></center>
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<center>Fig.2 The result of DNA sequencing  </center>
  
  
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===Usage and Biology===
 
  
 
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<partinfo>BBa_K5236013 parameters</partinfo>
 
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===Reference===
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Ding, Zundan, et al. “Rational Redesign of Thermophilic Pet Hydrolase LCCICCG to Enhance Hydrolysis of High Crystallinity Polyethylene Terephthalates.” Journal of Hazardous Materials, Elsevier, 7 Apr. 2023, www.sciencedirect.com/science/article/pii/S0304389423006696.

Latest revision as of 12:31, 2 October 2024

LCCICCG

Cutinase (EC 3.1.1.74) is a lipolytic/esterolytic enzyme that hydrolyzes not only cutin, which is a major component of plant cuticl, but also water-soluble esters and insoluble triglycerides. Leaf-branch compost cutinase (LCC) was first reported in 2012 and its activity and thermostability improved via four mutations yielding LCC-ICCG.

Usage and Biology

We tested for successful plasmid construction and transformation into E.coli through colony PCR and gel electrophoresis. The following gel result demonstrates that the plasmid transformed into E.coli are correct. The plasmid should have a total of 891 base pairs and the results match.


Fig.1 The DNA gel electrophoresis result

Fig.2 The result of DNA sequencing


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 258
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 258
    Illegal NheI site found at 190
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 258
    Illegal XhoI site found at 775
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 258
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 258
  • 1000
    COMPATIBLE WITH RFC[1000]



Reference

Ding, Zundan, et al. “Rational Redesign of Thermophilic Pet Hydrolase LCCICCG to Enhance Hydrolysis of High Crystallinity Polyethylene Terephthalates.” Journal of Hazardous Materials, Elsevier, 7 Apr. 2023, www.sciencedirect.com/science/article/pii/S0304389423006696.