Composite

Part:BBa_K1885456

Designed by: Kien Patrick Malarney   Group: iGEM16_ASIJ_Tokyo   (2016-10-29)


Novel osmY-PETase fusion protein, A1.00

PETase is a novel enzyme for the degradation of polyethylene terephthalate (PET) plastic. It was discovered in Ideonella sakaiensis, a bacteria species found by screening PET-exposed microbe communities in Japan. It functions as the first step in the degradation of PET; PETase catalyzes the hydrolysis of PET into terephthalate and ethylene glycol monomers. The N-terminal osmY protein and its signal peptide fused to PETase, with serine-glycine peptide repeats intervening, serves to secrete the PETase out of the cell so that it can act on PET in the cell's local environment. The Anderson promoter with a relative strength of 1.00 that precedes the fusion protein serves to constitutively express the fusion PETase, enhancing the degradation of environmental PET by the presence of more fusion PETase by constitutive expression. This part offers a more efficient initial step for teams interested in the bioremediation of PET, because the first step involved in PET bioremediation is hydrolysis into products that can then be utilized by a cell. Teams can then metabolically engineer bacteria of interest to metabolize ethylene glycol and terephthalate, but initially, these monomers must be produced by hydrolysis. Thus, PET bioremediation depends on the production of metabolizable monomers, and this part offers more efficient production of such monomers.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1543
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal PstI site found at 1543
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1367
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1543
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1543
    Illegal NgoMIV site found at 818
    Illegal NgoMIV site found at 872
    Illegal NgoMIV site found at 899
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
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