"
Team:UC Davis/Project/Catalyst
From 2012.igem.org
(Difference between revisions)
| Line 1,243: | Line 1,243: | ||
list-style-type:none;color:#000000 "><li>Who we are </li><li>Students </li><li>Advisors</li></ul></a> </li> | list-style-type:none;color:#000000 "><li>Who we are </li><li>Students </li><li>Advisors</li></ul></a> </li> | ||
| - | <li style="float:left ;margin:0 10px;"><a href="https://2012.igem.org/Team:UC_Davis/Project "><p>Project</p></a> <ul style="text-indent:-15px;list-style-image:none;list-style-type:none;color:#000000"><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Project">Project Overview</a> </li><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Project/ | + | <li style="float:left ;margin:0 10px;"><a href="https://2012.igem.org/Team:UC_Davis/Project "><p>Project</p></a> <ul style="text-indent:-15px;list-style-image:none;list-style-type:none;color:#000000"><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Project">Project Overview</a> </li><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Project/Catalyst ">Module Engineering</a></li><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Project/Strain ">Chassis Engineering</a></li> |
| - | <li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Project/Directed_Evolution "> - Directed Evolution</a></li> | + | <li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Project/Directed_Evolution "> - Directed Evolution</a></li> |
| - | <li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Project/Our_Strain "> - Rational Engineering</a></li> | + | <li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Project/Our_Strain "> - Rational Engineering</a></li> |
| - | <li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Project/Protein_Engineering ">Protein Engineering</a></li><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Parts ">Parts</a></li> </ul> </li> | + | <li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Project/Protein_Engineering ">Protein Engineering</a></li><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Parts ">Parts</a></li> |
| + | <li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Criteria">Critera</a> </li></ul> </li> | ||
<li style="float:left ;margin:0 10px"><a href="https://2012.igem.org/Team:UC_Davis/Safety "> <p>Safety</p></a> <a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Safety "> Safety</a> </li> | <li style="float:left ;margin:0 10px"><a href="https://2012.igem.org/Team:UC_Davis/Safety "> <p>Safety</p></a> <a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Safety "> Safety</a> </li> | ||
| - | <li style="float:left ;margin:0 10px;"><a href="https://2012.igem.org/Team:UC_Davis/Notebook "> <p>Notebook</p></a> <ul style="text-indent:-15px;list-style-image:none;list-style-type:none;color:#000000 "><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Notebook/Overview">Overview</a> </li><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Notebook/Protocols ">Protocols</a> </li><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Notebook ">Notebook</a> </li><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Notebook/Gallery">Gallery</a> </li> </ul> </li> | + | <li style="float:left ;margin:0 10px;"><a href="https://2012.igem.org/Team:UC_Davis/Notebook/Overview "> <p>Notebook</p></a> <ul style="text-indent:-15px;list-style-image:none;list-style-type:none;color:#000000 "><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Notebook/Overview">Overview</a> </li><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Notebook/Protocols ">Protocols</a> </li><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Notebook">Notebook</a> </li><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Notebook/Gallery">Gallery</a> </li><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Criteria">Critera</a> </li> </ul> </li> |
| - | <li style="float:left ;margin:0 10px;"><a href="https://2012.igem.org/Team:UC_Davis/Data "> <p>Data </p>< | + | <li style="float:left ;margin:0 10px;"><a href="https://2012.igem.org/Team:UC_Davis/Data "> <p>Data </p></a> <a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Data "> Data</a> </li> |
| - | + | ||
<li style="float:left ;margin:0 10px"><a href="https://2012.igem.org/Team:UC_Davis/Attributions "> <p>Attribution </p></a><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Attributions "> Attribution</a></li> | <li style="float:left ;margin:0 10px"><a href="https://2012.igem.org/Team:UC_Davis/Attributions "> <p>Attribution </p></a><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Attributions "> Attribution</a></li> | ||
Revision as of 05:08, 30 September 2012
UCDavis iGEM Tweets
Our Sponsors
Modules
LC-Cutinase and Initial PET Degradation
We had the LC-Cutinase gene synthesized with a pelB leader sequence and a 6-his tag and labeled it Bba_K936014. It has been placed the following construct where it is promoted by a pBad variant Bba_K206000.
The pelB leader sequence on the cutinase gene directs the enzyme to the periplasmic space [1] . Once the enzyme is led towards the membrane, there is a leakage that helps it be secreted into the extracellular matrix [1]. We hoped that this sequence would help the enzyme be secreted so the PET would more easily be degraded. When we ordered the cutinase sequence, we added pelB to the front of the sequence, in hopes of repeating the secretion shown in previous results. The inclusion of a his-tag allows us to purify the cutinase protein and identify where it is after it is produced. We have designed and conducted an experiment to determine how much of the protein is being secreted and how much is remaining inside of the cell, the results of which can be found here (link coming soon).
Ethylene Glycol Modules
Glycolaldehyde reductase (DL-1,2-propanediol oxidoreductase) is normally an anaerobic protein that reduces L-lactaldehyde into L-1,2-propanediol, an excreted product. In addition, a mutant from the paper aforementioned was able to live on L-1,2-propanediol as a sole carbon source. Not only are there mutants that live on the L-1,2-propanediol, but there are also mutants selected for growth on ethylene glycol. We are working to mutate the reductase to work aerobically, rather than anaerobically.
In contrast to the reductase, glycolaldehyde dehydrogenase is an aerobic protein that oxidizes glycolaldehyde further to glycolate. The glycolate will be used further downstream in metabolism to provide the carbon source for the E. coli to live.
We are using these enzymes polycistronically with the Bba_J23101 and Bba_K206000 promoters to see the difference in overproduction of the enzymes and modulating the production. Our modular efforts in plasmids will eventually be applied toward a rational strain engineering approach, where we manipulate the MG1655 chromosome to optimize the degradation of ethylene glycol.
References
2. Boronat, Albert, Caballero, Estrella, and Juan Aguilar. “Experimental Evolution of a Metabolic Pathway for Ethylene Glycol Utilization by Escherichia coli.” Journal of Bacteriology, Vol. 153 No. 1, pp. 134-139, January 1983.
3. Lu, Zhe, Elisa Cabiscol, Nuria Obradors, Jordi Tamarit, Joaquim Ros, Juan Aguilar, and E.C.C. Lin. "Evolution of an Escherichia coli Protein with Increased Resistance to Oxidative Stress." Journal of Biological Chemistry. 273.14 (1998): n. page. Print.
