Composite

Part:BBa_K1582023

Designed by: Dongqi Bao   Group: iGEM15_Tianjin   (2015-09-05)

FsC+inJanus-m Fusion Protein


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 379
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 525
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 301
    Illegal BsaI site found at 841


Usage

This fusion protein is designed to degrade PET into terephthalic acid and ethylene glycol more powerfully. Compared to single FsC, the rate of PET emzymolysis is supposed to be improved by LC-Janus fusion.

Biology

FsC is a kind of cutinase, the details of which can be seen at BBa_K808025. Hydrophobin inJanus, from the edible mushroom Grifola frondosa, play a role in a broad range of processes in the growth and development of filamentous fungi. Their assembly could show rod-like structure. They can be expressed in eucaryotic cells. We did some mutations to it, in order to make it expressed in E.coli, and we call it inJanus-m.
According to some research, when Janus is added to the reaction of cutinase and plastics, the reaction rate is supposed to have an obvious improvement.
Based on the above background, we design to use the method of fusion protein to combine cutinase and Janus compactly, through the establishment of fusion protein, we can found if we can further promote the stimulation rate of the plastics degradation. Meanwhile, from the data about fusion protein, we can try to uncover the functional mechanism of the stimulation to the plastics degradation which is unknown until now.


References

[1]Zefang Wang, Shuren Feng, Yujian Huang, Shan Li, Haijin Xu, Xiuming Zhang, Yanling Bai, Mingqiang Qiao, Expression and characterization of a Grifola frondosa hydrophobin in Pichia pastoris, Protein Expression and Purification 72 (2010) 19–25.
[2]Doris Ribitsch, Enrique Herrero Acero, Agnes Przylucka, Enhanced cutinase-catalyzed hydrolysis of polyethylene terephthalate 2 by covalent fusion to hydrophobins, Appl. Environ. Microbiol. doi:10.1128/AEM.04111-14.

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