Part:BBa_K4221008
mPETase-GSlinker-BslA(42-181aa)
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 783
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 783
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 783
- 25INCOMPATIBLE 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 - 1000COMPATIBLE 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.
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
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.
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