Difference between revisions of "Part:BBa K4390072"
 
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'''This part is not compatible with BioBrick RFC10 assembly but is compatible with the iGEM Type IIS Part standard [[Help:Standards/Assembly/Type_IIS|which is also accepted by iGEM.]]'''
 
'''This part is not compatible with BioBrick RFC10 assembly but is compatible with the iGEM Type IIS Part standard [[Help:Standards/Assembly/Type_IIS|which is also accepted by iGEM.]]'''
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'''This is a Level 0 part of type O part generating the following 4 base overhangs at upstream (AGCC) and downstream (TTCG) ends.'''
  
 
==Usage and Biology==
 
==Usage and Biology==

Latest revision as of 23:56, 13 October 2022


WT-RolA Hydrophobin


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.

This is a Level 0 part of type O part generating the following 4 base overhangs at upstream (AGCC) and downstream (TTCG) ends.

Usage and Biology

RolA is 151 amino acid protein, belonging to a group of hydrophobins that are found in fungi. Hydrophobins can increase the hydrophilicity of the PET product surface, thereby increasing the affinity of the substrate for PETase and thus the release of the final product. Although the exact molecular mechanism of this method has not been fully elucidated, several published articles have demonstrated that such a treatment can increase the release of the final product (Puspitasari, Tsai and Lee, 2020) (Puspitasari, Tsai and Lee, 2021).

However, the special requirements of the working environment of hydrophobins need to be further addressed before they can be used. Hydrophobins are classified as Class I and Class II. Class I is only soluble in strong acids and bases such as trifluoroacetic acid or formic acid, while Class II is less extreme and can be dissolved in ethanol or sodium dodecyl sulphate, or by applying pressure or lowering the temperature (Puspitasari, Tsai and Lee, 2021).

Design

The part was designed as JUMP assembly O part, and the codon was optimized for BioBrick and JUMP assembly. However, the lack of LacO between promoter and RBS should be fixed by future teams to achieve inducible expression of the Soluble RolA.

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 57
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 49

References

Puspitasari N, Tsai S, Lee C. Class I hydrophobins pretreatment stimulates PETase for monomers recycling of waste PETs. International Journal of Biological Macromolecules. 2021;176:157-164.

Puspitasari, N., Tsai, S. and Lee, C., 2020. Fungal Hydrophobin RolA Enhanced PETase Hydrolysis of Polyethylene Terephthalate. Applied Biochemistry and Biotechnology, 193(5), pp.1284-1295.