(7 intermediate revisions by 3 users not shown) | |||
Line 1: | Line 1: | ||
− | + | ||
{{HK_GTC}} | {{HK_GTC}} | ||
<html> | <html> | ||
+ | |||
+ | <style> | ||
+ | .section-heading{ | ||
+ | background-image: url("https://static.igem.org/mediawiki/2021/4/4d/T--HK_GTC--res2.jpg"); | ||
+ | } | ||
+ | </style> | ||
+ | |||
+ | <span class="material-icons to-top"><a href="#the-top">expand_less</a></span> | ||
+ | |||
+ | <div class="section-heading" id="the-top"> | ||
+ | <div class="mask"></div> | ||
+ | <h1>Contribution</h1> | ||
+ | </div> | ||
+ | |||
+ | <div class="section-wrapper"> | ||
+ | |||
+ | <div class="section-contents-mid large-img"> | ||
+ | <p>In the existing part BBa_K2982005, we added Scanning Electron Microscope (SEM) photos of PET films after | ||
+ | being digested by S245I PETase mutant. The usage of SEM can accurately visualize how serious the PET film is | ||
+ | digested by the enzyme. Besides, we added HPLC profile of products released from PET film digested with | ||
+ | S245I PETase to show the extent of PET hydrolysis. | ||
+ | |||
+ | </p> | ||
+ | |||
+ | <h2>(A) SEM images of digested PET films</h2> | ||
+ | <p>We performed PET film digestion to study the PET hydrolytic activity of S245I PETase. After incubating 9μg | ||
+ | of purified proteins with PET film at 30°C for 96 hours, we viewed the digested PET film under a SEM. | ||
+ | </p> | ||
+ | |||
+ | <div class="single-image-with-desc"> | ||
+ | <center><img src="https://static.igem.org/mediawiki/2021/c/ce/T--HK_GTC--contribution_1.png" alt=""></center> | ||
+ | </div> | ||
+ | |||
+ | <p>The pitting resulting from the digestion of S245I PETase mutant can be visualized, and the buffer-only SEM | ||
+ | photo acts as a control. | ||
+ | |||
+ | </p> | ||
+ | |||
+ | <p>(B) HPLC profile of the products released from the PET films</p> | ||
+ | |||
+ | <div class="single-image-with-desc"> | ||
+ | <center><img src="https://static.igem.org/mediawiki/2021/c/ce/T--HK_GTC--Contribution--a.png" alt=""></center> | ||
+ | <p>Figure 2a. HPLC spectrum of TPA standard. </p> | ||
+ | </div> | ||
+ | |||
+ | <div class="single-image-with-desc"> | ||
+ | <center><img src="https://static.igem.org/mediawiki/2021/5/58/T--HK_GTC--2b.png" alt=""></center> | ||
+ | <p> Figure 2b. HPLC spectrum of MHET standard.</p> | ||
+ | </div> | ||
+ | |||
+ | <div class="single-image-with-desc"> | ||
+ | <center><img src="https://static.igem.org/mediawiki/2021/9/99/T--HK_GTC--2d.png" alt=""></center> | ||
+ | <p> Figure 2c. HPLC spectrum of products released from the PET film digested with S245I PETase.</p> | ||
+ | </div> | ||
+ | |||
+ | <p>HPLC profiles demonstrated that the detection peaks representing TPA monomer and MHET intermediate product | ||
+ | formed during PET film digestion have retention times at 4.64 minutes (Figure 2a) and 5.17 minutes (Figure | ||
+ | 2b) respectively. <br> | ||
+ | HPLC profiles of products released from PET film digestions using S245I PETase revealed incomplete PET | ||
+ | hydrolysis as considerable amounts of intermediate product, MHET which showed a peak at 5.07 minutes were | ||
+ | detected (Figure 2c). <br> | ||
+ | Therefore, we can conclude that S245I PETase exhibits hydrolytic activity for PET depolymerization. | ||
+ | </p> | ||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | + | </div> | |
− | + | </div> | |
− | + | ||
− | </ | + | |
− | + | ||
− | + | ||
− | |||
− | < | + | <div class="clear"></div> |
− | </ | + | <footer> |
− | </ | + | <div> |
+ | <h1>GT COLLEGE iGEM2021</h1> | ||
+ | <img width="30%" src="https://static.igem.org/mediawiki/2021/7/78/T--HK_GTC--logo.png" alt=""> | ||
+ | </div> | ||
+ | <div> | ||
+ | <h1>Follow Us!</h1> | ||
+ | <a href="https://www.instagram.com/gtigemteam/"> | ||
+ | <i class="icon fab fa-instagram fa-2x"></i> | ||
+ | </a> | ||
+ | </div> | ||
+ | <div> | ||
+ | <h1>Contact:</h1> | ||
+ | <p>igemteam.gt@gmail.com</p> | ||
+ | </div> | ||
+ | </footer> | ||
+ | </html> |
Latest revision as of 14:22, 20 October 2021