Difference between revisions of "Part:BBa K1921003"
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<partinfo>BBa_K1921003 parameters</partinfo>
 
<partinfo>BBa_K1921003 parameters</partinfo>
 
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===Usage===
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The PET-ase is an enzyme, which can hydrolyze PET. And this mutation protein is changed on the basis of the PET–ase. This protein is changed from R to T at 64 position. This mutation protein can hydrolyze PET in low temperature, which is a big problem in the world. It also can increase in the bacteria, which is easy to get.
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===Biology===
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Compared with the sequence of PETase in NCBI, the homology of PETase and Enzyme TfCut2 was 52%. The paper says that scientists Ren Wei found that change TfCut2 enzyme amino acid 63 T into other amino acids, the degradation products of the reaction will inhibit the reaction, and the vicinity amino acid sequence at the site of TfCut2 enzyme has a high homology with PETase. Based on the literatures, we decided to mutate the amino acid R at this position to T for lower inhibition from product.
 +
 +
===Protein Expression===
 +
The bacteria were cultured in 5mL LB liquid medium with ampicillin in 37℃ 5h. After taking samples, we used 1mM IPTG 5μl induced in 16℃ for 4-5h. Then we make the expression .

Revision as of 17:22, 1 October 2016


PETase MutateM


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]


Usage

The PET-ase is an enzyme, which can hydrolyze PET. And this mutation protein is changed on the basis of the PET–ase. This protein is changed from R to T at 64 position. This mutation protein can hydrolyze PET in low temperature, which is a big problem in the world. It also can increase in the bacteria, which is easy to get.

Biology

Compared with the sequence of PETase in NCBI, the homology of PETase and Enzyme TfCut2 was 52%. The paper says that scientists Ren Wei found that change TfCut2 enzyme amino acid 63 T into other amino acids, the degradation products of the reaction will inhibit the reaction, and the vicinity amino acid sequence at the site of TfCut2 enzyme has a high homology with PETase. Based on the literatures, we decided to mutate the amino acid R at this position to T for lower inhibition from product.

Protein Expression

The bacteria were cultured in 5mL LB liquid medium with ampicillin in 37℃ 5h. After taking samples, we used 1mM IPTG 5μl induced in 16℃ for 4-5h. Then we make the expression .