Team:NCTU Formosa/Project

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{| style="color:#1b2c8a;background-color:rgb(255,178,60);" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"
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<p id="back-top"><a href="#top"><span></span>Back to Top</a></p>
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!align="center"|[[Team:NCTU_Formosa|Home]]
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!align="center"|[[Team:NCTU_Formosa/Team|Team]]
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!align="center"|[https://igem.org/Team.cgi?year=2012&team_name=NCTU_Formosa Official Team Profile]
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!align="center"|[[Team:NCTU_Formosa/Project|Project]]
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      <h1><a href="https://2012.igem.org/Team:NCTU_Formosa/Project">Introduction</a></h1>
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!align="center"|[[Team:NCTU_Formosa/Parts|Parts Submitted to the Registry]]
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      <a href="https://2012.igem.org/Team:NCTU_Formosa/Project"><p>Introduction</p></a>
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!align="center"|[[Team:NCTU_Formosa/Modeling|Modeling]]
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!align="center"|[[Team:NCTU_Formosa/Notebook|Notebook]]
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!align="center"|[[Team:NCTU_Formosa/Safety|Safety]]
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      <h1><a href="https://2012.igem.org/Team:NCTU_Formosa/Project-sub2#sub2-1">Enzyme for Isobutanol</a></h1>
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!align="center"|[[Team:NCTU_Formosa/Attributions|Attributions]]
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      <a href="https://2012.igem.org/Team:NCTU_Formosa/Project-sub2#sub2-1"><p>Enzyme for Isobutanol</p></a>
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      <h1><a href="https://2012.igem.org/Team:NCTU_Formosa/Project-sub2#sub2-2">Temperature Control System</a></h1>
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      <a href="https://2012.igem.org/Team:NCTU_Formosa/Project-sub2#sub2-2"><p>Temperature Control System</p></a>
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== '''Overall project''' ==
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      <h1><a href="https://2012.igem.org/Team:NCTU_Formosa/Project-sub2#sub2-3">Zinc Finger</a></h1>
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      <a href="https://2012.igem.org/Team:NCTU_Formosa/Project-sub2#sub2-3"><p>Zinc Finger</p></a>
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Our team invented an enzyme pathway for E. coli to produce butanol, which is closely related chemically to ethanol. However, the two extra carbon atoms and their attendant hydrogens let it be a higher energy value. In terms of storage and handling, it is less corrosive than ethanol and has several other advantages. Therefore, it may be a better substitution to ethanol as a biofuel.<br><br>
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The pathway starts at glucose, through glycolysis it will convert to pyruvate. Then the pyruvate goes through enzymatic process and produces isobutyraldehyde, which is toxic to the E.coli. To prevent the toxicity harming E.coli, we use the temperature regulation system.
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      <h1><a href="https://2012.igem.org/Team:NCTU_Formosa/Project-sub2#sub2-4">Instrument</a></h1>
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The first part is [[The 37℃ part]]. Our team makes E. coli produce a lot of nontoxic 2-Ketoisovalerate. Afterwards we reduce the temperature to 30℃.[[The 30℃ part ]] converts 2-ketoisovalerate to isobutyraldehyde at one blast. Then through simple process we get isobutanol, the potential biofuel in the future.<br><br>
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      <a href="https://2012.igem.org/Team:NCTU_Formosa/Project-sub2#sub2-4"><p>Instrument</p></a>
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What worth mentioning is that our team adds various [[zinc fingers]] before each enzyme to increase the affinity and efficiency of gene expression. Also, we are planning to design a container to produce and concentrate isobutanol. [[Team:NCTU_Formosa|NCTU_Formosa]] hopes to soon provide an affordable and “green” transportation fuel to the world!<br><br>
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== '''Project Details''' ==
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      <h1><a href="https://2012.igem.org/Team:NCTU_Formosa/Project-sub3">Conclusion</a></h1>
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The process of our experiment can be summarized by the following process.
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      <a href="https://2012.igem.org/Team:NCTU_Formosa/Project-sub3"><p>Conclusion</p></a>
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[[image:pathway.png]]
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      <h1><a href="https://2012.igem.org/Team:NCTU_Formosa/Project-sub4">Optimization</a></h1>
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Because one of the products of our experiment is harmful to the E.coli ,we want to resolve the problem. If the problem did’t disappear ,we would let E.coli be degraded. Then we decided to use 37℃ system to regulate our E.coli.<br><br>
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      <a href="https://2012.igem.org/Team:NCTU_Formosa/Project-sub4"><p>Optimization</p></a>
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[[image:main 37CS.png]]<br><br>
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First ,we make E.coli stay in 37℃ environment. E.coli will produce tetR to inhibit the production of isobutyraldehyde which is toxic to E.coli.<br><br>
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      <h1><a href="https://2012.igem.org/Team:NCTU_Formosa/Project-sub5">Future Works</a></h1>
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[[image:37CC.png]]<br><br>
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      <a href="https://2012.igem.org/Team:NCTU_Formosa/Project-sub5"><p>Future Works</p></a>
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After all of 2-ketoisovalerate are produced completely , we begin to let E.coli stay in 30℃ environment. The change will make E.coli begin the next reaction. It can be showed by the following process.<br><br>
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[[image:37CCC.png]]<br><br>
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<h1 id="project-s1-title" class="project-s-title"><a name="sub1"> </a>　<span>Introduction to the project</span></h1>
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<p>Nowadays, environmental pollution and energy depletion have become crucial problems. We need to find alternative energy to replace the running out fossil fuel. Due to the pollution issues, this alternative energy should be environmental friendly. Up until now, ethanol is the most common biomass fuel because the final product is harmless water. However, ethanol will corrode metallic  surface of the engines lead to higher cost than fossil fuel usage. Unlike ethanol, isobutanol do not corrode metal and contain higher ratio of the heat of combustion than ethanol. Besides, as well as ethanol, isobutanol doesn’t produce pollutants such as sulfur dioxide, nitric oxide and nitric dioxide. Isobutanol has widely utilized in many applications as a organic solvent, and antifreeze. As what we want, to get clean energy, we chose isobutanol to be our project. We believe that isobutanol is a potential eco fuel in the future. However, currently isobutanol production wasn't very promising. According to the previous studies, the low yield of isobutanol was caused by the toxicity of isobutanol which would kill the host <i>E.coli</i> . In this study, we introduced two innovative and brilliant solutions to solve this serious problem. Now, let’s take a deeper look in our new ideas!</p></div>
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Finally, we can obtain the main product, isobutanol, from the reaction.Our resolution is to promote all 2-ketoisovalerate turn into the toxic intermediate at a time for decreasing the strength of E.coli inhibition from isobutyaldehyde.<br><br>
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<div id="counter"><img src="https://static.igem.org/mediawiki/2012/d/d1/NctuCapture.jpg" /><img src="http://s11.flagcounter.com/count/Rhz/bg_FFFFFF/txt_000000/border_CCCCCC/columns_2/maxflags_248/viewers_3/labels_1/pageviews_1/flags_1/" /><img src="https://static.igem.org/mediawiki/2012/d/d1/NctuCapture.jpg" /></div>
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==='''Result'''===
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    <p>Copyright © 2012 NCTU Formosa</p>
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<p>Background image belongs to <a href="http://fatezoom.deviantart.com/" target="_blank">Yuri Kormin</a>.</p>
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== '''ZINC FINGER''' ==
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    <p class="author">Website designed by Calvin Hue.</p>
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<b>A zinc finger is a kind of finger-shaped protein that contains one or more zinc ions to stabilize the structure. Proteins that contain zinc fingers(zinc finger protein) are the most common transcription factors in organisms. A transcription factor is a protein that binds to DNA and controls the expression of genes. The zinc finger proteins bind specific DNA, RNA, or other molecules with the interaction of its domain and these molecules.<br><br>
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We add zif268,PBSII,HIVC and Gli1 four zinc fingers separately in front of our biopart: Alss, ilvc, ilvD, kivd. We expect that the enzyme with zinc fingers would increase the productivity of isobutanol.<br><br>
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=== zif268 ===
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[[image:Zif268.jpg|thumb|Zif268 Binding With DNA]]
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This  zinc finger identifies the DNA sequence 5'-GCG TGG GCG-3',binding to the certain domain of the sequence.<br><br><br><br><br><br><br><br><br><br><br><br>
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=== PBSII  ===
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Identifies DNA sequence 5'-GTG TGG AAA-3'.<br><br><br><br><br><br><br><br><br><br>
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=== HivC  ===
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Identifies DNA sequence 5'-GAT GCT GCA-3'.<br><br><br><br><br><br><br><br>
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=== Gli1  ===
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[[image:2GLI.pdb1.jpg|thumb|Five-Finger GLI/DNA Complex]]
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Identifies DNA sequence 5'-GAC CAC CCA-3'.<br><br><br><br><br><br><br><br>
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Latest revision as of 15:15, 26 October 2012

Team:NCTU Formosa - 2012.igem.org

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　Introduction to the project

Nowadays, environmental pollution and energy depletion have become crucial problems. We need to find alternative energy to replace the running out fossil fuel. Due to the pollution issues, this alternative energy should be environmental friendly. Up until now, ethanol is the most common biomass fuel because the final product is harmless water. However, ethanol will corrode metallic surface of the engines lead to higher cost than fossil fuel usage. Unlike ethanol, isobutanol do not corrode metal and contain higher ratio of the heat of combustion than ethanol. Besides, as well as ethanol, isobutanol doesn’t produce pollutants such as sulfur dioxide, nitric oxide and nitric dioxide. Isobutanol has widely utilized in many applications as a organic solvent, and antifreeze. As what we want, to get clean energy, we chose isobutanol to be our project. We believe that isobutanol is a potential eco fuel in the future. However, currently isobutanol production wasn't very promising. According to the previous studies, the low yield of isobutanol was caused by the toxicity of isobutanol which would kill the host E.coli . In this study, we introduced two innovative and brilliant solutions to solve this serious problem. Now, let’s take a deeper look in our new ideas!

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