Composite

Part:BBa_K4390093

Designed by: Zhongyi Liang   Group: iGEM22_Edinburgh-UHAS_Ghana   (2022-09-14)


His-tagged Soluble 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

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.

The soluble version RolA with 6His-tag for Nickel affinity column purification. The wild-type RolA forms insoluble assemblies, so this soluble version of RolA was used to see how this property would affect PETase activity.


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]

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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