Difference between revisions of "Part:BBa K4221008"

 
 
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===Usage and Biology===
 
  
 
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===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. 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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===Biology===
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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.
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BslA is a structurally defined bacterial hydrophobin that was found in the biofilm of Bacillus subtilis.
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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]
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===Design Consideration===
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The construct was cloned into a pET28a plasmid and transformed into BL21 (DE3) E. coli.
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===References===
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[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
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<!-- Add more about the biology of this part here
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===Usage and 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