This year, we hold couples of forums in order to talk with great biology professors about our project. According to the ardent brainstorm, we proposed a advanced theory and built a proof of concept system called Training Protein System. All the work we did was centered on validating whether the system is applicable to all enzymes or not. Training Protein System is a universalizable system for all the protein enzymes. Its main idea is a kind of research ideas, which is the most unique thing of it. Specifically, first and the foremost, researchers should analyze the protein tertiary structure and find its enzyme active center. The next step has two paths to improve the activity of enzymes. One was to mutate its gene sequence; the other was to make a surface display separately using E.coli and Yeast. We demonstrate it working under real condition. This year, we use the system on PETase to ensure if this system can apply in real practice. What is particularly noteworthy is we used the hydrophobic protein to modify the surface of reaction system to increase its activity. It is really lucky that we managed to have a satisfactory result in training PETase to work better.
The degradation of normal plastics
To find a better way to solve the problem of white pollution, we went to PET factory for a visit to learn more about PET and its industry. In factory, we know the latest technology of PET. Before we leaving, we get some PET samples from the director of the factory. It really interest us result in that we only use the standardized PET samples which are only used in labs for degradation and HPLC detection. This time we can do that on the normal plastic which we called hcPET in order to show our system working under real word conditions. Our HPLC results is shown as following.
Figure 1. Relative enzyme activity of engineering bacteria E.coli(BL21)/pET22b(+)LAP when they degarade PET bottles in different contitions.
Exploring for the best conditions by modeling
In order to apply the tech in the project into reality, we design a software which can assist us in processing data of HPLC. Our software can calculate peak area of any substance by the raw data of HPLC conveniently and accurately. It can display the image details saved at any period of time. It also can automatically identify substance peak and calculate the area. In addition, we can manually select the range of the area calculated. What’s more, in order to facilitate the analysis, it can store the results of calculations in a same file which contains the filename of the HPLC data. Our software can be used on Windows system. The installation manual is attached to our software. And we have a user guide to explain how to operate our software conveniently. Meanwhile, we built a multivariate nonlinear regression analysis model to predict the optimum condition of recombination cell decomposing PET. We set three variables as the reaction conditions in our model, which respectively were the cell concentration, the time and temperature of protein-induced expression. Then we established a multivariate nonlinear regression equation after the reasonable analysis. Finally, we calculated the optimal condition of PET decomposition which means we can make the degradation of the PET much more efficient.