Composite parts we have designed
BBa_K1951004 : DesA producer
Part Composition:
This part is composed of 3 subparts :
- RBS: Ribosome Binding Site
- pARA/araC inductible promoter
- Lysine decarboxylase
Lysine decarboxylase DesA:
Lysine descarboxylase from Streptomyces coelicolor is an enzyme from the lyase family that converts lysine to cadaverin. The enzyme realizes the carbonyl group of the lysin amino acid. Cadaverine(1,5-diaminopentane) is a primary diamine which alkaline environment. The lysine decarboxylase is an enzyme induced the synthesis of which is promoted by anaerobiosis and an acidic pH.
In bacteriology, this enzyme is sought through the middle of Moeller lysine or medium lysine Taylor.
We registered the original sequence of this subpart in the iGEM registry of standard parts (BBa_K1951000). We optimized our sequence for E.coli and ordered the synthesis by addition of an inductible promoter.
BBa_K1941008 : FliC E. coli producer
Part Composition:
This part is a composite part composed of 2 Biobricks :
- BBa_K1951005: flagelln C
- BBa_K808007: TPA Transporter
NB-Esterase:
This enzyme catalyses the degradation of PET into Terephthalic acid (TPA) and Ethylene Glycol. It is the first and slowest reaction of our degradation chain. Indeed, it takes two weeks to degrade PET.
We performed an enzymatic assay of NB-Esterase using 4-nitrophenylbutyrate as a substrate because it has a close chemical structure to PET and we figured that this enzyme digested pNP in our bacteria
TPA Transporter:
The NB-Esterase is present in the extracellular medium and cuts PET in this location. In order to be fully degraded, the TPA needs to be integrated to our chassis organism which is why we incorporated a transporter to our degradation operon.
Part Assembly:
The subparts were assembled using standard BioBrick Assembly.
