Composite

Part:BBa_K4390092

Designed by: Zhongyi Liang   Group: iGEM22_Edinburgh-UHAS_Ghana   (2022-09-14)
Revision as of 00:35, 14 October 2022 by Dianajorge (Talk | contribs)

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His-tagged WT RolA

This part is not compatible with BioBrick RFC10 assembly but is compatible with the iGEM Type IIS Part standard which is also accepted by iGEM.

Usage and Biology

The wild-type RolA with 6His-tag for Nickel affinity column purification. RolA is part of a class of small fungal proteins called hydrophobins (Puspitasari, et al. 2021). As an 11kDa protein, it has been shown to increase PET hydrolysis, when in the form of a recombinant RolA-PETase. RolA is a highly surface-active substance that assembles spontaneously at hydrophobic-hydrophilic interfaces. By doing so, it is thought that the presence of RolA on PET surfaces makes the hydrophilic PETase able to readily hydrolyse the PET.


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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 169
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 161

Reference

Puspitasari, N., Tsai, SL. & Lee, CK. Fungal Hydrophobin RolA Enhanced PETase Hydrolysis of Polyethylene Terephthalate. Appl Biochem Biotechnol 193, 1284–1295 (2021). https://doi.org/10.1007/s12010-020-03358-y


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