Difference between revisions of "Part:BBa K4221008"
 
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===Usage===
 
===Usage===
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PET hydrolase (PETase), which hydrolyzes polyethylene terephthalate (PET) into soluble building blocks, provides an attractive avenue for the bioconversion of plastics.  
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PET hydrolase (PETase), which hydrolyzes polyethylene terephthalate (PET) into soluble building blocks, provides an attractive avenue for the bioconversion of plastics. mPETase is a complicated mutation of PETase.Our team used the amphiphilicity of BslA to enhance PET degrading efficiency of degrading enzyme mPETase.
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In the process of protein purification by ATPs, we can use the amphiphilicity of BslA to change the hydrophilicity of fluorescent protein, so that fluorescent protein can only show fluorescence in the organic phase/aqueous phase, so as to achieve a high-efficiency and low-cost protein purification method. Our team used the amphiphilicity of BslA to enhance the antibacterial/targeting effect of LL37 antimicrobial peptide and RGD-targeted peptide
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===Biology===
 
===Biology===

Latest revision as of 06:39, 11 October 2022


mPETase-GSlinker-BslA(42-181aa)

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 783
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 783
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 783
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 783
    Illegal NgoMIV site found at 58
    Illegal NgoMIV site found at 112
    Illegal NgoMIV site found at 139
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage

PET hydrolase (PETase), which hydrolyzes polyethylene terephthalate (PET) into soluble building blocks, provides an attractive avenue for the bioconversion of plastics. mPETase is a complicated mutation of PETase.Our team used the amphiphilicity of BslA to enhance PET degrading efficiency of degrading enzyme mPETase.

Biology

PETase is a plastic degrading enzyme derived from Ideonella sakaiensis, mPETase is a complicated mutation of PETase, which contains 11 mutation sites: S214H-I168R-W159H-S188Q-R280A-A180I-G165A-Q119Y-L117F-T140D-S121E.

BslA is a structurally defined bacterial hydrophobin that was found in the biofilm of Bacillus subtilis. It helps the assembling of TasA (an exopolysaccharide and an amyloid fiber-forming protein), the component of the biofilm matrix. BslA is composed of an Ig-type fold with the addition of an unusual, extremely hydrophobic “cap” region. The central hydrophobic residues of the cap are essential to allow a hydrophobic, nonwetting biofilm to form as they control the surface activity of the BslA protein.[1]

Design Consideration

The construct was cloned into a pET28a plasmid and transformed into BL21 (DE3) E. coli.

References

[1]: “BslA is a self-assembling bacterial hydrophobin that coats the Bacillus subtilis biofilm.” Proceedings of the National Academy of Sciences of the United States of America vol. 110,33 (2013): 13600-5. doi:10.1073/pnas.1306390110