Part:BBa_K3288062
light inducible expression of signal peptide (NSP4) fused MHETase(18-604;W397A)
BBa_K3288062 is a composite part in which BBa_K1075044, BBa_K3288007 and BBa_K3288017 are combined.
This part is compatible with RFC10.
Usage and Biology
After PETase and MHETase was analyzed based on protein structures, some of them were reported to be point mutated with increased enzyme activation. Our team checked PETase wild type, W159H/S238F, and S121E/D186H/R280A’s PET degradation activities to choose enzymes for PET degrading device. These enzymes were produced at 18 °C in LB (buffered 50mM BTP (pH8.5) for 24 hours. After centrifuged, we put PET disc into cleared media with no E.coli and reacted it for 24 hours at 40 degrees Celsius. Everyday, we replaced media with ones that has new PET degrading enzyme secreted. After 5 days, we washed PET disc and dried it up to measure the change in weight. Results showed that NSP4-PETase (S121E/D186H/R280A) degraded the most PET.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 2171
Illegal BamHI site found at 4015 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 63
Illegal NgoMIV site found at 195
Illegal NgoMIV site found at 289
Illegal NgoMIV site found at 582
Illegal NgoMIV site found at 1076
Illegal NgoMIV site found at 1094
Illegal NgoMIV site found at 1184
Illegal AgeI site found at 414
Illegal AgeI site found at 1542 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1643
Illegal BsaI site found at 3479
Illegal BsaI site found at 4283
Illegal BsaI.rc site found at 525
| None |