Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Tri Ekawati Heryanto, Joko Pebrianto Trinugroho, Pande Putu Erawijantari, Nimas Ghasani, Risma Wiharyanti, Gilang Tirta Widi, Oktira Roka Aji
Date created: 2014-09-28 11:00:00
Date modified: 2015-05-08 01:10:13
J23106 - tetO - B0034 - lpp ompA-LC-Cutinase - B1002
| Types | DnaRegion |
| Roles | engineered_region Composite |
| Sequences | BBa_K1387006_sequence (Version 1) |
Description
The casette of degradation module is comprised of constitutive promoter, tet operator, RBS, outer membrane protein A (ompA) fused with LC Cutinase, and double terminator. By using constitutive promoter, we expect the recombinant protein, in this case ompA-LC Cutinase, will be synthesized indefinitely. ompA is outer membrane protein. It comprises of β???strand B3-B7. Rather than using full sequence of ompA, we use ompA (46-159) fused with lipoprotein signal sequence, thus exposing it on the cell surface of bacteria1. We fuse lpp-ompA with LC Cutinase, an enzyme that exhibit esterase activity. LC-Cutinase esterase activity will cleave the ester bond on PET and releasing terephthalic acid and ethylene glycol as a product. Fusion strategy of lpp-ompA on LC Cutinase gene is one of our way to create a whole cell biocatalyst which will be an effective molecular machinery to degrade PET on the environment. Due to constitutive expression of the recombinant protein in E.coli, there is a possibility of formation of insoluble protein inside the cell (inclusion body), because of that, we put tet operator upstream of RBS for further regulation.Notes
Fusion strategy of lpp-ompA on LC Cutinase gene is one of our way to create a whole cell biocatalystSource
This part, we desgned through synthetic gene (G-Blocks)| Access | Instance | Definition |
| public public public public public public | BBa_J23106 BBa_K079036 BBa_B0034 BBa_K103006 BBa_K936014 BBa_B1002 | BBa_J23106 BBa_K079036 BBa_B0034 OmpA BBa_K936014 BBa_B1002 |
| Sequence Annotation | Location | Component / Role(s) |
| BBa_J23106 BBa_K079036 BBa_B0034 BBa_K103006 BBa_K936014 BBa_B1002 | 1,35 44,58 67,78 87,550 557,1525 1534,1567 | BBa_J23106 BBa_K079036 BBa_B0034 OmpA BBa_K936014 BBa_B1002 |