Team:AHUT-ZJU-China/Contribution
AHUT-ZJU-China
Contribution
On the basis of OT3-CA-WT, we tried several mutations by means of molecular simulation in our project, and finally identified our OT3-CA-MU. In the course of this project, we have gained a lot, and made useful contributions to the future iGEM teams. We will list our contributions in detail below:
1.We measured the enzyme activity of OT3-CA-WT and compared its enzyme activity at high temperature and low temperature. By measuring the enzyme activity, the future iGEMers can see the change of wild-type CA enzyme activity more intuitively, which is convenient for them to determine the maximum enzyme activity. Through determination, iGEM team can obtain more direct changes of wild type enzyme activity in future. Also they can determine the temperature corresponding to maximum activity of enzyme activity.
2.We measured the enzyme activity of OT3-CA-WT at different temperatures. Through our data, future IGEM teams could more directly see the changes in the activity of OT3-CA-WT , so that they could determine the corresponding temperature when the enzyme activity value reached the maximum.
Fig.1 Induced expression of OT3-CA-WT in TB1 strain verified by SDS-PAGE
Fig. 2 Induced expression of OT3-CA-WT in TB1 strain verified by Western blot
Fig. 3 Esterase activity analysis of OT3-CA-WT protein at different temperatures
3.We have identified the DNA sequence of the OT3-CA-MU, which we have entered into the Parts and can be accessed and used directly by future iGEM teams.
4.We tried to compare the human carbonic anhydrase (CA2) with the mutation of position 203 in 2018 and our OT3-CA-WT this year. By changing the sequence of human carbonic anhydrase (CA2) on the basis of the original, we hope that if we can further improve the thermal stability of human beings.
Fig.4 A schematic diagram of the structure of human carbonic anhydrase amino acids after mutation at position 203.
5.Our project can be studied as a scientific research project. Greenhouse Effect is a concern of all mankind, and CO2 is the main cause of greenhouse effect, so reducing CO2 is a fundamental solution to this problem. The follow-up of our project will become a topic to continue studying how to better use the methods of synthetic biology to capture CO2, and our team will endeavor to make contributions to the humanity.