Results


Results of SDS-PAGE

After performing SDS-PAGE of our protein complex, our result finally came out. However, to be honest, our result isn't as ideal as we imagine at first. The following picture is our result of SDS-PAGE. As the photo shows, all of the proteins have more or less inaccuracy except BBa_K4618823. We would like to analyze our result below.

1. BBa_K4618823: The most distinct band occurred between 25 and 35. Our result showed a very close data compared with the ideal protein size. Therefore, we considered BBa_K4618823 to have a quite accurate data.
2. BBa_K4618500, BBa_K4618501,BBa_K4618502, BBa_K4618213: these four laccase didn't have an obvious band. Therefore, we cannot read their protein sizes at first. We tried to improve their sequence to make the distinct band appear.
After our improvement, we perform the experiment again. This time, there are some distinct bands appeared. However, the bands weren't precisely matched with our ideal protein sizes, which means that our experiment still have a long way to go. We would try to perfect our experiment in the future. The following photo is our second result, and we would like to analyze them below.

1. BBa_K4618500:Though not really distinct, a band occurred at about 60, but the protein size is 76.01, not really match with our result.
2. BBa_K4618501:It show an obvious band at about 25, which is far away from our ideal result. Finding a way to revise our protein is our important future work .
3. BBa_K4618502:We still cannot read the band easily, we are guessing whether there is any external interference we didn't think of that made our result inaccurate.
4. BBa_K4618105: The distinct band occurred between 75 and 100, which matched with the ideal protein size. It was the second protein complex we design to match with our experiment.
Though our improvement cannot make all the protein complex perfectly match with the ideal protein size, we do our best to purify the protein complex, and the result became much closer to what we predicted.



Laccase Measure Result

Due to our 3D model built In dry lab, we found that BBa_K4618105 cannot perfectly bind with laccase. Thus, we only use laccase to perform the color-degrading experiment, and measure BBa_K4618105 binding with color catcher individually. After our final experiment, our result finally came out. The following form is our result.



Analysis


1. BBa_K4618500:it perform well in acid dyes(yellow and blue), but doesn't have an obvious change in disperse dye(red) and azo dye(black), we thought these two kinds of dyes may have some kinds of ways to prevent the laccase from degrading colors.
2. BBa_K4618501:This kind of protein didn't perform any degrading behaviors. We thought we may have made some mistakes during the protein purification. It would be our future work to find out our mistake.
3. BBa_K4618502:This one can degrade all kinds of color, we thought that this kind of protein can fit well in acidic environment, which is our dyes made, leading it to a very successful result.
4. BBa_K4618213:This one only cannot make the disperse dye degraded, we thought the disperse dye may be providing an uncomfortable environment for BBa_K4618213, inhibits it from being active.

Future Plan

1. We will try more ways to purify our protein, make them match with our ideal protein sizes.
2. We will make researches about why BBa_K4618501 cannot work as we imagine, and try to improve it in the future.
3. We will analyze the relationship between the working efficiency of the protein and the characteristic of the dye, and try to synthesize a protein complex that can fit the environment made by dyes.

Consideration for replicating the experiment

We would like to add some salt in dye water to make it become buffer solution, and provide a better environment to stay active.