Difference between revisions of "Part:BBa K2910002"
 
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<partinfo>BBa_K2910002 parameters</partinfo>
 
<partinfo>BBa_K2910002 parameters</partinfo>
 
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In order to assess the enzymatic activity of PETase variants relative to the wild type, and W159H S238F PETase (BBa_k2910000), p-nitrophenyl butyrate assays were performed. Ester bonds in PET plastic are mimicked by p-nitrophenol. Each PETase variant cleaves the ester bond; once the bond is broken it emits a wavelength at 405 nm. This emission can be read by a plate reader; if PETase is catalytically active, absorbance at 405 should be perceived. The following figures show the results of the aforementioned assay for this variant.
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A plot of the pNB Hydrolysis at pH = 8 by BBa_K2910002 is presented below:
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<img src="https://parts.igem.org/File:T--Toronto--pNPB-Hydrolysis-pH-8_Tianyu-All-Conc.png">
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A plot of the pNB Hydrolysis at pH = 9 by BBa_K2910002 is presented below:
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<img src="https://parts.igem.org/File:T--Toronto--pNPB-Hydrolysis-pH-9_Tianyu-All-Conc.png">

Latest revision as of 03:58, 22 October 2019


IsPETase-ML Mut 1 with C-terminal Hexahistidine Tag

BBa_K2910002 encodes a variant of the Ideonella sakaiensis PETase, optimized for E. coli, which degrades poly(ethylene terephthalate. Using a Machine Learning Algorithm to optimize hydrophobicity content, the thermostability of the protein is predicted to be improved.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]



In order to assess the enzymatic activity of PETase variants relative to the wild type, and W159H S238F PETase (BBa_k2910000), p-nitrophenyl butyrate assays were performed. Ester bonds in PET plastic are mimicked by p-nitrophenol. Each PETase variant cleaves the ester bond; once the bond is broken it emits a wavelength at 405 nm. This emission can be read by a plate reader; if PETase is catalytically active, absorbance at 405 should be perceived. The following figures show the results of the aforementioned assay for this variant.

A plot of the pNB Hydrolysis at pH = 8 by BBa_K2910002 is presented below: <img src="https://parts.igem.org/File:T--Toronto--pNPB-Hydrolysis-pH-8_Tianyu-All-Conc.png">

A plot of the pNB Hydrolysis at pH = 9 by BBa_K2910002 is presented below: <img src="https://parts.igem.org/File:T--Toronto--pNPB-Hydrolysis-pH-9_Tianyu-All-Conc.png">