Team:TUST China/Improve
Improve Parts
| Name | Type | Description | Designer | Length | |
 |
BBa_K2685000 | Coding | Lip | LIN,WENXIN | 1113 |
|
BBa_K2685001 | Coding | Mnp | LIN,WENXIN | 1131 |
 |
BBa_K2685016 | Composite | pTet+Lip+LacI+pLac+φX174E | LIN,WENXIN | 2723 |
This year we selected lignin peroxidase (Lip) and manganese peroxidase (Mnp), which are non-specific lignin-structure decomposing enzymes produced by white rot fungi. It can effectively degrade PCBs, PAHs, DDT, fuels, explosives, other chlorides and azides in wastewater and soil.
This year we selected the lignin peroxidase (BBa_K500000) and manganese peroxidase(BBa_K500001) from iGEM10_Tianjin in 2010. They are expressed in yeast and can oxidate phenol type which are rich in electronic or non phenolic aromatic compounds, since the electron transport system can attack substrates. After that, it can take an electron from phenol or benzene rings of phenols to oxide subsrances into free radicals, which in turn to produce many different kinds of free radical chain reactions, thereby tetracycline.can be degraded and eliminated.
This year we selected the lignin peroxidase (BBa_K500000) and manganese peroxidase (BBa_K500001) from iGEM10_Tianjin in 2010. They are expressed in yeast and can oxidate phenol type which are rich in electronic or non phenolic aromatic compounds, since the electron transport system can attack substrates. After that, it can take an electron from phenol or benzene rings of phenols to oxide subsrances into free radicals, which in turn to produce many different kinds of free radical chain reactions, thereby tetracycline.can be degraded and eliminated.
However, in our preliminary experiments, we found that they could hardly be expressed in E. coli.
Fig.1 the growth curve of the E.coli with BBa_K500000:and empty plasmid under tetracycline environment
Therefore, we optimized the codon to make it be highly expressed in E. coli. After verification, we got the following graph which indicated that the parts we constructured with optimized codons can work productively and efficiently. Thus, we use these two enzymes as a benchmark to construct a part: BBa_K2685011 Part: BBa_K2685012 Part: BBa_K2685013. Finally, we found that lip enzyme has the best and the most desirable effect of degradation compared with other parts.
Fig.2 the growth curve of the E.coli with BBa_K2685011:and BBa_K500000 under tetracycline environment