Parts

Overview

In our project, we have divided it into three main components:

1. Dye Removal:
        In this first part, we have employed four different Laccases produced by various species. Laccase enzymes are selected for their ability to break down benzene rings, thus achieving the desired color removal effect.

2. Integration with Color Catcher:
        To enable the binding of Laccase enzymes to Color Catcher, we have utilized hydrophobin. By modifying the surface properties of the Color Catcher, hydrophobin facilitates the binding process.

3. Protein Linkage:
        Finally, the most crucial aspect of our project involves connecting all the proteins. We have utilized the NG system to create a bridge between laccase and hydrophobin, ensuring their effective linkage.

Parts collection

Part improvement

In order to enable laccase to better bind to the PET paper, we wanted to combine it with hydrophobin. However, due to possible protein folding problems and sequence length issues, we cloned them into two plasmids separately. So we found a protein called NGcatcher/NGtag.

NGcatcher/tag is a system that combines two proteins through isopeptide bonds. And the BBa_K4618823 in it can cover the surface of NGcatcher and stabilize it.

During the process of designing the gene, we found that the proportion of GC in the BBa_K4618823 was too high, resulting in the gene being unable to be successfully synthesized. Therefore, we optimized the BBa_K4618823 to reduce the proportion of GC in its sequence without changing the protein.

Lastly, we connected BBa_K4618805, BBa_K4618213, BBa_K4618500 ,BBa_K4618501, and BBa_K4618502 so that we hope that they can be successfully expressed and combined with each other to generate functions.

Fig 1.Binding Reaction