Team:KULeuven/Project

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__NOTOC__
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''Our wiki has been frozen. For updates check [https://2009.igem.org/Main_Page iGEM's Main Page].
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''Follow our adventures at the Jamboree in Boston on our [http://igemkuleuven.wordpress.com/ BLOG].''
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<html><center></html>
 
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{| border="0" class="Generic"
 
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|-
 
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!
 
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! class="top" |  [[Team:KULeuven/Modelling|Modelling]]
 
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! class="top" |  Lab
 
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! class="top" |  Notebook
 
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! class="left" |
 
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|  [[Team:KULeuven/Notebook|General Notebook]]
 
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! class="left" | Blue Light Receptor
 
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|  [[Team:KULeuven/Modelling/Blue Light Receptor|Blue Light Receptor Modelling]]
 
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|  [[Team:KULeuven/Lab/Blue Light Receptor|Blue Light Receptor Lab]]
 
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|  [[Team:KULeuven/Notebook/Blue Light Receptor|Blue Light Receptor Notebook]]
 
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! class="left" |  Vanillin Production
 
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|  [[Team:KULeuven/Modelling/Vanillin Production|Vanillin Production Modelling]]
 
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|  [[Team:KULeuven/Lab/Vanillin Production|Vanillin Production Lab]]
 
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|  [[Team:KULeuven/Notebook/Vanillin Production|Vanillin Production Notebook]]
 
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! class="left" |  Vanillin Receptor
 
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|  [[Team:KULeuven/Modelling/Vanillin Receptor|Vanillin Receptor Modelling]]
 
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|  [[Team:KULeuven/Lab/Vanillin Receptor|Vanillin Receptor Lab]]
 
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|  [[Team:KULeuven/Notebook/Vanillin Receptor|Vanillin Receptor Notebook]]
 
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! class="left" |  Key/Lock/AntiKey
 
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|  [[Team:KULeuven/Modelling/Key/Lock/AntiKey|Key/Lock/AntiKey Modelling]]
 
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|  [[Team:KULeuven/Lab/Key/Lock/AntiKey|Key/Lock/AntiKey Lab]]
 
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|  [[Team:KULeuven/Notebook/Key/Lock/AntiKey|Key/Lock/AntiKey Notebook]]
 
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<div id="imap2wrapper">
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<div id="promovid"><object width="213" height="172"><param name="movie" value="http://www.youtube.com/v/TlfIiHTczsY&hl=en&fs=1&"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/TlfIiHTczsY&hl=en&fs=1&" type="application/x-shockwave-flash" allowscriptaccess="always" allowfullscreen="true" width="213" height="172"></embed></object></div>
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<div id="smallabstract">
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<a href="#Abstract">
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<p>
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<b><i>Essencia coli</i></b> is a <b>vanillin producing</b> bacterium equipped with a <b>control system</b> that keeps the concentration of vanillin at a constant level. Take a <b>guided tour</b> by clicking the different parts in the figure. <b>Click here for a project abstract.</b>
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<dt><a id="title" href="#" title="Factory Figure"></a></dt>
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<dd id="pic1"><a id="text1" title="Blue Light Receptor" href="https://2009.igem.org/Team:KULeuven/Design/Blue_Light_Receptor"><span></html>{{Team:KULeuven/Components/Blue_Light_Receptor}}<html></span></a></dd>
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<dd id="pic2"><a id="text2" title="Vanillin Receptor" href="https://2009.igem.org/Team:KULeuven/Design/Vanillin_Receptor"><span></html>{{Team:KULeuven/Components/Vanillin_Receptor}}<html></span></a></dd>
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<dd id="pic3"><a id="text3" title="Key Lock Antikey" href="https://2009.igem.org/Team:KULeuven/Design/Key_Antikey"><span></html>{{Team:KULeuven/Components/Key_Lock_Antikey}}<html></span></a></dd>
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<dd id="pic4"><a id="text4" title="Vanillin Production" href="https://2009.igem.org/Team:KULeuven/Design/Vanillin_Production"><span></html>{{Team:KULeuven/Components/Vanillin_Production}}<html></span></a></dd>
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<dd id="pic5"><a id="text5" title="CEO Introduction" href="#Miss_Blue_Vanilla:_CEO_of_the_Fragrance_Factory"></a></dd>
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==Miss Blue Vanilla: CEO of the Fragrance Factory==
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[[image:Miss_Blue+tekst.png|700px]]
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= Essencia Coli =
 
== Abstract ==
== Abstract ==
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=== Vanilla Perfume, the story of a control bacterium ===
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‘''Essencia coli''’ is a vanillin producing bacterium equipped with a control system that keeps the concentration of vanillin at a constant level. The showpiece of the project is the feedback mechanism. Vanillin synthesis is initiated by irradiation with blue light. The preferred concentration can be modulated using the intensity of that light. At the same time the bacterium measures the amount of vanillin outside the cell and controls its production to maintain the set point. The designed system is universal in nature and has therefore potential benefits in different areas. The concept can easily be applied to other flavours and odours. In fact, any application that requires a constant concentration of a molecular substance is possible.
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‘Essencia Coli’ is a vanillin producing bacterium equipped with a control system that keeps the concentration of vanillin at a constant level. The novelty of the project is the feedback mechanism. Vanillin synthesis is initiated by irradiation with blue light. The preferred concentration can be modulated using the intensity of that light. At the same time the bacterium measures the amount of vanillin outside the cell and controls its production to maintain the set point. An additional advantage lies in the universal nature of the system. The concept can easily be applied to other flavours and odours: any application that requires a constant concentration of a molecular substance.
 
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== concept  ==
 
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[[Image:Mssbluevanilla.jpg|right|thumb|425px|Schematic illustration of the setup for controlling concetration of molecule 'X']]
 
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The main idea is to synthesize Vanillin and control the synthesis such that the concentration of Vanillin stays constant. The value of this concentration can be set by intensity of blue light. This concept however can be extended to other molecules that need to be kept at a certain concentration, this can be for example other odors or flavours or something completely different like essential nutrients or vitamins. In the figure on the right an illustration of the controlling mechanism is explained. In here Vanillin is changed to molecule 'X' because of it's universal charachter. In the figure the cell synthesizes molecule 'X' and at the same time measures the concentration of this molecule outside the cell  and uses this measurement to control the synthesis and keep the concentration outside the cell constant. This concentration can then be modulated using light intensity.
 
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For proof of concept the molecule 'X' is set to be vanillin, to extend this principle to other molecule all that is needed is a concentration dependent receptor. The light receptor will activate transcription of A on irradiation of blue light. This will stimulate synthesis of Vanillin. The concentration vanillin outside the cell is measured with a vanillin receptor. The activation of the vanillin receptor will result in transcription of B and blocks the actions of A. The activation of synthesis by A is done with a key/lock setup. The translation of the Vanillin synthesizing proteins need a key in order to translate, A will provide the key. Therefore upon irradiation of blue light the required proteins will be translated. The inhibition of A by B is done by translating an anti-sense mRNA of A, blocking the actions of the key. Thus, if the concentration of vanillin increases, the synthesis of vanillin-synthesizing proteins will decrease untill it stops completely.
 
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The complete system with the corresponding biobricks can be seen below, as well as the biobricks which still need to be created from the genes (blue light receptor and vanillin synthesis)
 
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[[Image:Missbluevanilla.jpg|center|thumb|400px|Overview of the system with corresponding biobricks]]
 
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{{Team:KULeuven/Common2/PageFooter}}

Latest revision as of 20:41, 21 October 2009

Our wiki has been frozen. For updates check iGEM's Main Page. Follow our adventures at the Jamboree in Boston on our [http://igemkuleuven.wordpress.com/ BLOG].


Blue Light Receptor

The receptor senses the blue light to which the bacteria are exposed. By choosing the light intensity, the wanted vanillin concentration is set. Upon photo-excitation the receptorprotein YcgF dimerizes and interacts directly with the repressor YcgE. This protein is bound to the promoter-region of the key-gene and inhibits its transcription. The dimerized YcgF acts as an anti-repressor and releases YcgE from the DNA. Thereupon, the transcribed key activates vanillin production, which is proportional to the entered intensity of the blue light.

Vanillin Receptor

The vanillin receptor senses the vanillin concentration outside the cell. This information is needed to create the feedback loop that controls vanillin synthesis. The receptor consists of two proteins: virA and virG. In the presence of vanillin, virA binds a phosphor and transfers it to virG. In turn, the phosphorylated VirG binds to a so-called vir box sequence, triggering transcription of antikey. The more antikey is produced, the more it can anneal to the key and the less vanillin is produced. This process goes on until there’s an equilibrium between measured and wanted concentration.

Key Lock Antikey

The key/antikey system compares the signal from the blue light and vanillin receptor in order to control vanillin production. The more the measured amount of vanillin exceeds the wanted amount (set by the blue light intensity), the less vanillin is produced. After blue light irradiation, the key is transcribed and ‘unlocks’ the vanillin synthesis pathway. In response to the produced vanillin, the vanillin receptor then activates transcription of the antikey. Key and antikey are complementary RNA-strands: their annealing is favoured over the reaction between key en lock leading to vanillin synthesis.

Vanillin Production

A vanilla odour is created by synthesizing the molecule Vanillin. The starting point is tyrosine, an amino acid produced endogenously in E.coli. The subsequent pathway involves a combination of five enzymes. By locking both the transcription of the first and the third enzyme we prevent vanillin synthesis without the presence of the key.


Miss Blue Vanilla: CEO of the Fragrance Factory

Miss Blue+tekst.png

Abstract

‘Essencia coli’ is a vanillin producing bacterium equipped with a control system that keeps the concentration of vanillin at a constant level. The showpiece of the project is the feedback mechanism. Vanillin synthesis is initiated by irradiation with blue light. The preferred concentration can be modulated using the intensity of that light. At the same time the bacterium measures the amount of vanillin outside the cell and controls its production to maintain the set point. The designed system is universal in nature and has therefore potential benefits in different areas. The concept can easily be applied to other flavours and odours. In fact, any application that requires a constant concentration of a molecular substance is possible.




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