Difference between revisions of "Team:ITB Indonesia/Demonstrate"
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<h1 class="ITB_h1" style="padding-bottom: 30px; margin-bottom: 50px; border-bottom: 2px solid #1c2922 !important; padding-left: 30px; text-align: center; color: #1c2922">Demonstrate</h1> | <h1 class="ITB_h1" style="padding-bottom: 30px; margin-bottom: 50px; border-bottom: 2px solid #1c2922 !important; padding-left: 30px; text-align: center; color: #1c2922">Demonstrate</h1> | ||
| − | <p> | + | <p style="font-size: 18px"><strong>Real World's Conditions, Real World's Materials</strong></p> |
| − | <p> | + | <p>In order to demonstrate the work of our project in real world's conditions, we did not only use chemical test reagents (such as pNPB) to test our PETase activity. We instead tried to find the realest material that would be encountered by our bacteria on its applications. We ended up using plastic bottles directly as materials to be degraded.</p> |
| + | <p>The results of commonly found plastic bottle degradation by our bacteria are quite satisfactory. SEM analyses showed that our bacteria are able to degrade even real life plastic bottles, which are not pure PET. The effect is shown by SEM analyses where the plastic surface inoculated by our PETase-containing bacteria are rougher and has more cracks compared to its control.</p> | ||
| + | <center><img src="https://static.igem.org/mediawiki/parts/a/a0/T--ITB_Indonesia--SEMControl.jpeg" style="width: 500px; height: auto; img-align:center"></center> | ||
| + | <center><em> Figure 2. SEM result of plastic fragment incubated with control BL21 cells bearing no plasmid </em></center> | ||
| + | <br> | ||
| + | <center><img src="https://static.igem.org/mediawiki/parts/7/72/T--ITB_Indonesia--SEMPETase.jpeg" style="width: 500px; height: auto; img-align:center"></center> | ||
| + | <center><em> Figure 3. SEM result of plastic fragment incubated with BL21 cells transformed with BBa_K2378005 shows rougher surface with more cracks </em></center> | ||
| + | <br> | ||
| + | <center><img src="https://static.igem.org/mediawiki/parts/c/c2/T--ITB_Indonesia--SEMNhaR.jpeg" style="width: 500px; height: auto; img-align:center"></center> | ||
| + | <center><em> Figure 4. SEM result of plastic fragment incubated with BL21 cells transformed with BBa_K2378006 shows rougher surface with more cracks </em></center> | ||
| + | <br> | ||
| + | <p style="font-size: 18px"><strong>Real World's Conditions, Real World's Inducers</strong></p> | ||
| + | <p>Aside from using daily materials, we also tried to incorporate the use of real world's inducers. Our bacteria are supposed to be induced naturally when they encounter plastic, due to the presence of organic pollutants (OPs) on the surface of the plastic.</p> | ||
| + | <p>In order to mimic real world's conditions, we also tried to induce the expression of PETase in our bacteria through exposure with plastic wastes.</p> | ||
| + | <p>The culture induced by plastic wastes did not have the same expression rate as cultures induced with common chemical inducer salicylate. However, it turned out that some degree of expression is observed in cultures induced with plastic wastes, albeit the difference of the expression rate compared to uninduced cultures are not too significant.</p> | ||
| + | <center><img src="https://static.igem.org/mediawiki/parts/6/6b/T--ITB_Indonesia--psalpnpb.jpeg" style="width: 500px; height: auto; img-align:center"></center> | ||
| + | <center><em> Figure 5. pSal-Regulated PETase Activity (pNPB Assay) of BBa K2378004 Transformants</em></center> | ||
</div> | </div> | ||
Latest revision as of 03:26, 2 November 2017
Demonstrate
Demonstrate
Real World's Conditions, Real World's Materials
In order to demonstrate the work of our project in real world's conditions, we did not only use chemical test reagents (such as pNPB) to test our PETase activity. We instead tried to find the realest material that would be encountered by our bacteria on its applications. We ended up using plastic bottles directly as materials to be degraded.
The results of commonly found plastic bottle degradation by our bacteria are quite satisfactory. SEM analyses showed that our bacteria are able to degrade even real life plastic bottles, which are not pure PET. The effect is shown by SEM analyses where the plastic surface inoculated by our PETase-containing bacteria are rougher and has more cracks compared to its control.
Real World's Conditions, Real World's Inducers
Aside from using daily materials, we also tried to incorporate the use of real world's inducers. Our bacteria are supposed to be induced naturally when they encounter plastic, due to the presence of organic pollutants (OPs) on the surface of the plastic.
In order to mimic real world's conditions, we also tried to induce the expression of PETase in our bacteria through exposure with plastic wastes.
The culture induced by plastic wastes did not have the same expression rate as cultures induced with common chemical inducer salicylate. However, it turned out that some degree of expression is observed in cultures induced with plastic wastes, albeit the difference of the expression rate compared to uninduced cultures are not too significant.
