Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Kien Patrick Malarney
Date created: 2016-10-13 11:00:00
Date modified: 2016-10-29 11:17:00
Gibson-assembled pSB1C3 with novel fusion osmY-PETase A0.58
| Types | DnaRegion |
| Roles | engineered_region Composite |
| Sequences | BBa_K1885002_sequence (Version 1) |
Description
PETase is a novel enzyme for the degradation of polyethylene terephthalate (PET) plastic. It was discovered in Ideonella sakaiensis, a bacteria species found by screening PET-exposed microbe communities in Japan. It functions as the first step in the degradation of PET; PETase catalyzes the hydrolysis of PET into terephthalate and ethylene glycol monomers.The N-terminal osmY protein and its signal peptide fused to PETase, with serine-glycine peptide repeats intervening, serves to secrete the PETase out of the cell so that it can act on PET in the cell's local environment.
The Anderson promoter with a relative strength of 0.58 that precedes the fusion protein serves to constitutively express the fusion PETase, enhancing the degradation of environmental PET by the presence of more fusion PETase through constitutive expression.
This part offers a more efficient initial step for teams interested in the bioremediation of PET, because the first step involved in PET bioremediation is hydrolysis into products that can then be utilized by a cell. Teams can then metabolically engineer bacteria of interest to metabolize ethylene glycol and terephthalate, but initially, these monomers must be produced by hydrolysis. Thus, PET bioremediation depends on the production of metabolizable monomers, and this part offers more efficient production of such monomers.
This part utilizes the pSB1C3 backbone and confers resistance to chloramphenicol.
Notes
Designing this part required considering constitutive expression and promoter strength, secretion of the PETase, and maintaining an active PETase conformation. Respectively, the Anderson promoter, osmY, and serine-glycine linker peptide address the aforementioned considerations.Source
The pSB1C3 backbone, the Anderson promoter, and serine-glycine linker peptide parts came from the iGEM registry. The N-terminal osmY sequence came from the Escherischia Coli genome, found on NCBI, and the PETase protein came from the Ideonella sakaiensis genome, found on NCBI and annotated as ???lipase."| Access | Instance | Definition |
| public public | BBa_J18921 BBa_J23111 | 3xGS BBa_J23111 |
| Sequence Annotation | Location | Component / Role(s) |
| BBa_J18921 BBa_J23111 | 1,18 27,61 | 3xGS BBa_J23111 |