Team:Saint Joseph/Basic Part
Basic Parts
Basic Parts
The laccase enzyme was found in 1883 in a tree called Rhus Vernifica. Later, this enzyme was found in other plants, bacteria, insects, but mostly in fungis.
One of the most important fungi with laccase enzyme is Trametes versicolor.The laccase enzyme consists of 500 amino acids. Maintains its stability between 5-55 degrees and is water soluble.
The molecular structure of laccase, such as copper ions, acidic amino acids, more than basic amino acids, makes it effective in many different fields.
The reason why we use Trametes versicolor laccase is that it is the best decolorizing enzyme for textile dyes.
The use of laccase instead of chlorinated compounds during the bleaching process in papermaking is more favorable to the environment, because chlorinated compounds remove toxic substances when bleaching.
The Trametes versicolor laccase gene (Laccase (lcc1) BBa_K2962000) is an optimized sequence for expression in the E. coli organism. Characterized by team iGEM10_Tianjin (BBa_K500002) to avoid codon degeneracy, we innovated a new sequence. We designed all of our other parts according to this laccase.
PET26b (BBa_K2962001), is an expression vector used for expression of Laccase gene in E. coli SHuffle organism.
Dsb Fusion tag (BBa_K2962002), induce the formation of disulfide bonds in the E. coli organism. In this study, dsb-tagged laccase was used for increase enzyme activity.
Histidine tag (BBa_K2962003), used for easy identification and purification of the expressed laccase enzyme. The laccase gene was purified with Nickel-bound Histidine tags.
Pelb signaling sequence (BBa_K2962004) was used for the periplasmic localization of the laccase enzyme in E. coli. This is expected to increase the yield of purificated protein. Characterized by BBa_K1127003 by team iGEM13_York_UK. We innovated a new gene sequence which is compatible with our laccase. It means we used an appropriate PelB signal sequence for our laccase enzyme.
Shuffle T7 E. coli Strain (BBa_K2962005) used for expression of designed and amplified laccase genes.
Saint Joseph Basic Parts
| Name | Type | Description | Designer | Length |
|---|---|---|---|---|
| BBa_K2962000 | Coding | lcc1 gene codes laccase enzyme | Handan Kesim | 1575 |
| BBa_K2962001 | Plasmid_Backbone | PET26b | Handan Kesim | 5360 |
| BBa_K2962002 | Tag | jdsb fusion tag | Handan Kesim | 17 |
| BBa_K2962003 | Tag | histidine tag | Handan Kesim | 18 |
| BBa_K2962004 | Signalling | Signalling | Handan Kesim | 63 |
| BBa_K2962005 | Cell | SHuffle T7 E.coli strain | Handan Kesim | - |
References
- [1] Mohamed Achraf Blout. Développement de biocathodes pour biopiles enzymatiques utilisant la laccase. Chimie théorique et/ou physique. Université Pierre et Marie Curie - Paris VI, 2017. Français. ffNNT : 2017PA066249ff. fftel-01695524f