Team:UCL/Project/Biobricks

From 2014.igem.org

(Difference between revisions)
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     <li><a href="#view1">UCL iGEM 2014</a></li>
     <li><a href="#view1">UCL iGEM 2014</a></li>
     <li><a href="#view2">Azoreductase</a></li>
     <li><a href="#view2">Azoreductase</a></li>
-
    <li><a href="#view3">Azoreductase 1B6</a></li>
 
     <li><a href="#view3">Laccase</a></li>
     <li><a href="#view3">Laccase</a></li>
     <li><a href="#view4">Lignin Peroxidase</a></li>
     <li><a href="#view4">Lignin Peroxidase</a></li>
-
     <li><a href="#view5">BsDyP</a></li>
+
     <li><a href="#view5">Dye Decolourisng Peroxidases</a></li>
-
     <li><a href="#view6">PpDyP</a></li>
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     <li><a href="#view6">Octaprenyl Diphosphate Synthase</a></li>
 +
    <li><a href="#view7">Extracellular Nuclease</a></li>
</ul>
</ul>
<div class="tabcontents">
<div class="tabcontents">
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<!--- This is the second biobrick --->
<!--- This is the second biobrick --->
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<div id="view3"><div class="textTitle"><h4>Azoreductase 1B6 (BBa_K1336001)</h4></div><br>
+
<div id="view3"><div class="textTitle"><h4>Spore Coat Protein Laccase (BBa_K1336002)</h4></div><br>
 +
<!-- This is the biobrick image -->
 +
<a data-tip="true" class="top large" data-tip-content="This diagram explains the basic construct of the BioBrick, the only part that changes is the selected function itself; in this case attributed to BBa_K1336001." href="javascript:void(0)" style="width: 30%;float: right;margin-left:2%"><img src="https://static.igem.org/mediawiki/2014/5/5d/UCLBBAzo1b6.png" style="max-width: 100%;"></a>
 +
<!-- This is the main text. Anything in a <p>TEXT</p> is a paragraph and will be spaced appropriately-->
 +
<p>Another azoreductase that we will be using is isolated from <em>Pseudomonas aeruginosa</em>. It functions in the same way as Azoreductase R1 -  cleaving the azo bond - but it is intended to work complementary with it, in order to cover a wider spectrum of dyes more efficiently. <br><br>Like the previous azoreductase, this BioBrick will be constructed using PCR. A promoter and a ribosomal binding site (RBS) will then be added to create a functioning composite device. </p><br>
 +
<div class="textTitle"><h4>Laccase (BBa_K729006)</h4></div><br>
<!-- This is the biobrick image -->
<!-- This is the biobrick image -->
<a data-tip="true" class="top large" data-tip-content="This diagram explains the basic construct of the BioBrick, the only part that changes is the selected function itself; in this case attributed to BBa_K1336001." href="javascript:void(0)" style="width: 30%;float: right;margin-left:2%"><img src="https://static.igem.org/mediawiki/2014/5/5d/UCLBBAzo1b6.png" style="max-width: 100%;"></a>
<a data-tip="true" class="top large" data-tip-content="This diagram explains the basic construct of the BioBrick, the only part that changes is the selected function itself; in this case attributed to BBa_K1336001." href="javascript:void(0)" style="width: 30%;float: right;margin-left:2%"><img src="https://static.igem.org/mediawiki/2014/5/5d/UCLBBAzo1b6.png" style="max-width: 100%;"></a>
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<!--- This is the third biobrick --->
<!--- This is the third biobrick --->
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<div id="view4"><div class="textTitle"><h4><a name="BBa_K1336003"><span>Lignin Peroxidase (BBa_K1336003)</span></a></h4></div><br>
+
<div id="view4"><div class="textTitle"><h4>Lignin Peroxidase (BBa_K500000)</h4></div><br>
<a data-tip="true" class="top large" data-tip-content="This diagram explains the basic construct of the BioBrick, the only part that changes is the selected function itself; in this case attributed to BBa_K1336003." href="javascript:void(0)" style="width: 30%;float: right;margin-left:2%"><img src="https://static.igem.org/mediawiki/2014/7/78/UCLBBLigningperoxidase.png" style="max-width: 100%;"></a>
<a data-tip="true" class="top large" data-tip-content="This diagram explains the basic construct of the BioBrick, the only part that changes is the selected function itself; in this case attributed to BBa_K1336003." href="javascript:void(0)" style="width: 30%;float: right;margin-left:2%"><img src="https://static.igem.org/mediawiki/2014/7/78/UCLBBLigningperoxidase.png" style="max-width: 100%;"></a>
<!-- This is the main text. Anything in a <p>TEXT</p> is a paragraph and will be spaced appropriately-->
<!-- This is the main text. Anything in a <p>TEXT</p> is a paragraph and will be spaced appropriately-->
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<!--- This is the fourth biobrick --->
<!--- This is the fourth biobrick --->
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<div id="view5"><div class="textTitle"><h4><a name="BBa_K1336004"><span><em>Bacillus subtilis</em> dye-decolorizing peroxidase (BsDyP) (BBa_K1336004)</span></a></h4></div><br>
+
<div id="view5"><div class="textTitle"><h4><em>Bacillus subtilis</em> dye-decolorizing peroxidase (BsDyP) (BBa_K1336003)</h4></div><br>
<a data-tip="true" class="top large" data-tip-content="This diagram explains the basic construct of the BioBrick, the only part that changes is the selected function itself; in this case attributed to BBa_K1336004." href="javascript:void(0)" style="width: 30%;float: right;margin-left:2%"><img src="https://static.igem.org/mediawiki/2014/a/ad/UCLBBBsdyp.png" style="max-width: 100%;"></a>
<a data-tip="true" class="top large" data-tip-content="This diagram explains the basic construct of the BioBrick, the only part that changes is the selected function itself; in this case attributed to BBa_K1336004." href="javascript:void(0)" style="width: 30%;float: right;margin-left:2%"><img src="https://static.igem.org/mediawiki/2014/a/ad/UCLBBBsdyp.png" style="max-width: 100%;"></a>
<!-- This is the main text. Anything in a <p>TEXT</p> is a paragraph and will be spaced appropriately-->
<!-- This is the main text. Anything in a <p>TEXT</p> is a paragraph and will be spaced appropriately-->
<p>Found in <em>B. subtilis</em>, the physiological function of this newly discovered enzyme is still unclear, although it has shown effectiveness in degrading lignin and azo dyes, which makes it useful for us. It is not as effective as PpDyP for most compounds, but very efficient in degrading ABTS (2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid)). <br><br>The BioBrick will be constructed via PCR.</p><br><br><br><br>
<p>Found in <em>B. subtilis</em>, the physiological function of this newly discovered enzyme is still unclear, although it has shown effectiveness in degrading lignin and azo dyes, which makes it useful for us. It is not as effective as PpDyP for most compounds, but very efficient in degrading ABTS (2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid)). <br><br>The BioBrick will be constructed via PCR.</p><br><br><br><br>
 +
<div class="textTitle"><h4><a name="BBa_K1336004"><span><em>Pseudomonas putida</em> MET94 dye-decolorizing peroxidase (PpDyP) (BBa_K1336005)</span></a></h4></div><br>
 +
<a data-tip="true" class="top large" data-tip-content="This diagram explains the basic construct of the BioBrick, the only part that changes is the selected function itself; in this case attributed to BBa_K1336005." href="javascript:void(0)" style="width: 30%;float: right;margin-left:2%"><img src="https://static.igem.org/mediawiki/2014/9/9c/UCLBBPpdyp.png" style="max-width: 100%;"></a>
 +
<!-- This is the main text. Anything in a <p>TEXT</p> is a paragraph and will be spaced appropriately-->
 +
<p>This enzyme is found in <em>P. putida</em>. Although it is relatively novel, and has not yet been studied in detail, it seem to be an extremely versatile and powerful biocatalyst; it oxidises a wide <a data-tip="true" class="top large" data-tip-content="Such as azo dyes, anthraquinones, phenolic compounds, manganese or veratryl alcohol." href="javascript:void(0)"><b>variety of substrates</b></a> very efficiently. This will broaden the <a data-tip="true" class="top large" data-tip-content="Going further just azo dyes!" href="javascript:void(0)"><b>spectrum of action</b></a> of our decolourising device, and thus being able to degrade other toxic compounds typically found in industrial wastewaters. <br><br>This BioBrick will be constructed via PCR.</p><br>
</div>
</div>
<!--- This is the fifth biobrick --->
<!--- This is the fifth biobrick --->
-
<div id="view6"><div class="textTitle"><h4><a name="BBa_K1336005"><span><em>Pseudomonas putida</em> MET94 dye-decolorizing peroxidase (PpDyP) (BBa_K1336005)</span></a></h4></div><br>
+
<div id="view6"><div class="textTitle"><h4>Octaprenyl Diphosphate Synthase (ispB) (BBa_K1336005)</h4></div><br>
<a data-tip="true" class="top large" data-tip-content="This diagram explains the basic construct of the BioBrick, the only part that changes is the selected function itself; in this case attributed to BBa_K1336005." href="javascript:void(0)" style="width: 30%;float: right;margin-left:2%"><img src="https://static.igem.org/mediawiki/2014/9/9c/UCLBBPpdyp.png" style="max-width: 100%;"></a>
<a data-tip="true" class="top large" data-tip-content="This diagram explains the basic construct of the BioBrick, the only part that changes is the selected function itself; in this case attributed to BBa_K1336005." href="javascript:void(0)" style="width: 30%;float: right;margin-left:2%"><img src="https://static.igem.org/mediawiki/2014/9/9c/UCLBBPpdyp.png" style="max-width: 100%;"></a>
<!-- This is the main text. Anything in a <p>TEXT</p> is a paragraph and will be spaced appropriately-->
<!-- This is the main text. Anything in a <p>TEXT</p> is a paragraph and will be spaced appropriately-->
<p>This enzyme is found in <em>P. putida</em>. Although it is relatively novel, and has not yet been studied in detail, it seem to be an extremely versatile and powerful biocatalyst; it oxidises a wide <a data-tip="true" class="top large" data-tip-content="Such as azo dyes, anthraquinones, phenolic compounds, manganese or veratryl alcohol." href="javascript:void(0)"><b>variety of substrates</b></a> very efficiently. This will broaden the <a data-tip="true" class="top large" data-tip-content="Going further just azo dyes!" href="javascript:void(0)"><b>spectrum of action</b></a> of our decolourising device, and thus being able to degrade other toxic compounds typically found in industrial wastewaters. <br><br>This BioBrick will be constructed via PCR.</p><br>
<p>This enzyme is found in <em>P. putida</em>. Although it is relatively novel, and has not yet been studied in detail, it seem to be an extremely versatile and powerful biocatalyst; it oxidises a wide <a data-tip="true" class="top large" data-tip-content="Such as azo dyes, anthraquinones, phenolic compounds, manganese or veratryl alcohol." href="javascript:void(0)"><b>variety of substrates</b></a> very efficiently. This will broaden the <a data-tip="true" class="top large" data-tip-content="Going further just azo dyes!" href="javascript:void(0)"><b>spectrum of action</b></a> of our decolourising device, and thus being able to degrade other toxic compounds typically found in industrial wastewaters. <br><br>This BioBrick will be constructed via PCR.</p><br>
</div>
</div>
 +
 +
<!--- This is the fifth biobrick --->
 +
<div id="view7"><div class="textTitle"><h4>Extracellular Nuclease (nucB) (BBa_K729004)</h4></div><br>
 +
<a data-tip="true" class="top large" data-tip-content="This diagram explains the basic construct of the BioBrick, the only part that changes is the selected function itself; in this case attributed to BBa_K1336005." href="javascript:void(0)" style="width: 30%;float: right;margin-left:2%"><img src="https://static.igem.org/mediawiki/2014/9/9c/UCLBBPpdyp.png" style="max-width: 100%;"></a>
 +
<!-- This is the main text. Anything in a <p>TEXT</p> is a paragraph and will be spaced appropriately-->
 +
<p>This enzyme is found in <em>P. putida</em>. Although it is relatively novel, and has not yet been studied in detail, it seem to be an extremely versatile and powerful biocatalyst; it oxidises a wide <a data-tip="true" class="top large" data-tip-content="Such as azo dyes, anthraquinones, phenolic compounds, manganese or veratryl alcohol." href="javascript:void(0)"><b>variety of substrates</b></a> very efficiently. This will broaden the <a data-tip="true" class="top large" data-tip-content="Going further just azo dyes!" href="javascript:void(0)"><b>spectrum of action</b></a> of our decolourising device, and thus being able to degrade other toxic compounds typically found in industrial wastewaters. <br><br>This BioBrick will be constructed via PCR.</p><br>
 +
</div>
 +
</div>
</div>

Revision as of 15:19, 16 October 2014

Goodbye Azodye UCL iGEM 2014

BioBricks

Our BioBricks & how they lead to azo degradation


We plan to create a complete synthetic azo dye decolourising device in E. coli which incorporates several different independent enzymes that act on azo dyes and their breakdown products. After evaluating their individual breakdown characteristics, we aim to investigate the potential synergistic action of these enzymes in a single synthetic E. coli device and design a bioprocess which could be used to upscale the method to an industrial context.


In an industrial setting, these enzymes would work sequentially in a bioreactor with preset dynamic conditions. First, azoreductase will cleave the azo-bond (N=N), producing a series of highly toxic aromatic amines. Then, these compounds will be oxidised by lignin peroxidase, laccase and bacterial peroxidases, completing decolourisation and decreasing toxicity levels.

The complementary action of azoreductase, lignin peroxidase, laccase, and bacterial peroxidases will be studied in order to find out the best possible approach of sequential reaction, and this core degradation module will be extrapolated to other areas such as BioArt projects and work on algal-bacterial symbiosis.


Azoreductase (BBa_K1336000)


This non-specific enzyme was isolated from Bacillus subtilis, although it is also found in other bacterial species. It starts the degradation of azo dyes by cleaving the azo bond.

The products of this cleavage varies greatly among different dyes, but are generally aromatic amines. This azo cleavage does not only occur with azo dyes, but also with other molecules like Sulfasalazine. We will isolate this enzyme from B. subtilis and convert it to BioBrick format via polymerase chain reaction (PCR).



Azoreductase 1B6 (BBa_K1336001)


Another azoreductase that we will be using is isolated from Pseudomonas aeruginosa. It functions in the same way as Azoreductase R1 - cleaving the azo bond - but it is intended to work complementary with it, in order to cover a wider spectrum of dyes more efficiently.

Like the previous azoreductase, this BioBrick will be constructed using PCR. A promoter and a ribosomal binding site (RBS) will then be added to create a functioning composite device.


Spore Coat Protein Laccase (BBa_K1336002)


Another azoreductase that we will be using is isolated from Pseudomonas aeruginosa. It functions in the same way as Azoreductase R1 - cleaving the azo bond - but it is intended to work complementary with it, in order to cover a wider spectrum of dyes more efficiently.

Like the previous azoreductase, this BioBrick will be constructed using PCR. A promoter and a ribosomal binding site (RBS) will then be added to create a functioning composite device.


Laccase (BBa_K729006)


Another azoreductase that we will be using is isolated from Pseudomonas aeruginosa. It functions in the same way as Azoreductase R1 - cleaving the azo bond - but it is intended to work complementary with it, in order to cover a wider spectrum of dyes more efficiently.

Like the previous azoreductase, this BioBrick will be constructed using PCR. A promoter and a ribosomal binding site (RBS) will then be added to create a functioning composite device.


Lignin Peroxidase (BBa_K500000)


Usually found in white-rot fungi species, its main function in nature is to participate in lignin-degrading processes by these organisms. However, it has also been found to play a role in azo dye degradation and decolourisation.

This enzyme, like laccase, would be incorporated in the second step of the reaction to oxidise the products of the azo bond cleavage, in order to achieve greater detoxification. The sequence for the enzyme will be ordered and synthesised, including the BioBrick prefix and suffix. Again, it will function together with a promoter and a RBS.


Bacillus subtilis dye-decolorizing peroxidase (BsDyP) (BBa_K1336003)


Found in B. subtilis, the physiological function of this newly discovered enzyme is still unclear, although it has shown effectiveness in degrading lignin and azo dyes, which makes it useful for us. It is not as effective as PpDyP for most compounds, but very efficient in degrading ABTS (2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid)).

The BioBrick will be constructed via PCR.






This enzyme is found in P. putida. Although it is relatively novel, and has not yet been studied in detail, it seem to be an extremely versatile and powerful biocatalyst; it oxidises a wide variety of substrates very efficiently. This will broaden the spectrum of action of our decolourising device, and thus being able to degrade other toxic compounds typically found in industrial wastewaters.

This BioBrick will be constructed via PCR.


Octaprenyl Diphosphate Synthase (ispB) (BBa_K1336005)


This enzyme is found in P. putida. Although it is relatively novel, and has not yet been studied in detail, it seem to be an extremely versatile and powerful biocatalyst; it oxidises a wide variety of substrates very efficiently. This will broaden the spectrum of action of our decolourising device, and thus being able to degrade other toxic compounds typically found in industrial wastewaters.

This BioBrick will be constructed via PCR.


Extracellular Nuclease (nucB) (BBa_K729004)


This enzyme is found in P. putida. Although it is relatively novel, and has not yet been studied in detail, it seem to be an extremely versatile and powerful biocatalyst; it oxidises a wide variety of substrates very efficiently. This will broaden the spectrum of action of our decolourising device, and thus being able to degrade other toxic compounds typically found in industrial wastewaters.

This BioBrick will be constructed via PCR.


Contact Us

University College London
Gower Street - London
WC1E 6BT
Biochemical Engineering Department
Phone: +44 (0)20 7679 2000
Email: ucligem2014@gmail.com

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