|
|
Line 252: |
Line 252: |
| <div class="two-columns block-text medium-sized not-centered no-margin-top" style="margin-bottom:0px"> | | <div class="two-columns block-text medium-sized not-centered no-margin-top" style="margin-bottom:0px"> |
| <div> | | <div> |
− | <p>
| |
− | But measurements of YFP in <i>C. reinhardtii</i> turned out to be a great challenge, because of the strong interaction of the algae with light (photo systems, pigments, chlorophyll and light antennae). More information on our process of measuring YFP can be found on our <a href="https://2019.igem.org/Team:Humboldt_Berlin/Measurement">Measurement page</a>.
| |
− | </p>
| |
− | </br>
| |
| <p> | | <p> |
| The overall goal of the model is to determine the time needed to degrade 1 mg of PET. The model took into account enzyme expression, secretion and kinetics and also the cultivation density of the algae as decisive factors for the PET degradation rate. The model predicted that for a cultivation density of 1:10, a 40 g PET bottle would be degraded in approximately 10 years. For a cultivation density of 1:100, the predicted time to degrade a bottle was 100 years. Additionally, an improvement of the PETase enzyme by a factor 1000 was made. For a cultivation density of 1:10 and the improved enzyme, the time needed to degrade a bottle was 119 days. For a cultivation density of 1:100 and the improved enzyme, the time needed to degrade a bottle was 3,3 years. The results of the model led us to take several decisions regarding the improvement of the PET degradation. We chose to use the light-inducible PsaD promoter for our constructs, the secretion enhancing glycomodule SP<sub>20</sub>, the specialized <i>C. reinhardtii</i> strain UVM4 for transgene expression and the flat panel cultivation method to achieve higher cultivation densities. For more information please visit our model page <a href="https://2019.igem.org/Team:Humboldt_Berlin/Model">here</a>. | | The overall goal of the model is to determine the time needed to degrade 1 mg of PET. The model took into account enzyme expression, secretion and kinetics and also the cultivation density of the algae as decisive factors for the PET degradation rate. The model predicted that for a cultivation density of 1:10, a 40 g PET bottle would be degraded in approximately 10 years. For a cultivation density of 1:100, the predicted time to degrade a bottle was 100 years. Additionally, an improvement of the PETase enzyme by a factor 1000 was made. For a cultivation density of 1:10 and the improved enzyme, the time needed to degrade a bottle was 119 days. For a cultivation density of 1:100 and the improved enzyme, the time needed to degrade a bottle was 3,3 years. The results of the model led us to take several decisions regarding the improvement of the PET degradation. We chose to use the light-inducible PsaD promoter for our constructs, the secretion enhancing glycomodule SP<sub>20</sub>, the specialized <i>C. reinhardtii</i> strain UVM4 for transgene expression and the flat panel cultivation method to achieve higher cultivation densities. For more information please visit our model page <a href="https://2019.igem.org/Team:Humboldt_Berlin/Model">here</a>. |