Difference between revisions of "Team:SSTi-SZGD/Description"

 
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<a href="https://2017.igem.org/Team:SSTi-SZGD/Applied_Design">Design</a>
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<span>Guardian of the wheatland</span>
 
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<span>A product for the degradation of soil pesticide residues</span>
 
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<p class="title">Pesticide Residue Sweeper--Why We Need it?</p>
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<p>
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The level of pesticide usage in China is 2.5 times above the world average. In June 2016, the total output of chemical pesticides in China was around 3.347 million tons, up by 7.17% compared with the same period of 2015. The deterioration effects of long-term and overdose usage of pesticide in soil include soil erosion, soil contamination, water pollution, organic pollution, reducing biodiversity, etc.
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<br />
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A range of chemical and physical methods have been applied to degrade pesticide residues. The shortfalls of these methods include high in cost, harmful to non-target organisms, food, plants and soils, and likely to cause secondary pollution. We intend to develop a microbial degradation method with low toxicity, low cost and high efficiency to serve a positive role in maintaining ecological balance.
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SSTi-iGEM combined optogenetics and biotechnology, using genetically modified E.coli as a carrier, to improve the microbial method for pesticide residue degradation in soil. We used a light-regulated gene expression system (LightOFF) that efficiently over-express heterogenous hydrolases which degrade insecticide organophosphorus or fungicide carbendazim. This application can result to two forms of products: enzyme and whole cell products. In addition, we propose to develop an automatic praying system that facilitates the application of whole cell products in real-life scenario.
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The level of pesticide usage in China is 2.5 times above the world average. In June 2016, the total output of chemical pesticides in China was around 3.347 million tons, up by 7.17% compared with the same period of 2015. The deterioration effects of long-term and overdose usage of pesticide in soil include soil erosion, soil contamination, water pollution, organic pollution, reducing biodiversity, etc. A range of chemical and physical technologies have been widely applied in the degradation of pesticide residues. These techniques are of high in accuracy and easy to be manipulated characteristics. However, the shortfalls come with high in cost, harmful to non-target organisms, food, plants and soils, and likely to cause secondary pollution, etc.
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<p class="title">How to do it?----Project Design</p>
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This year we aimed to to combine optogenetics and biotechnology, by using genetically modified organism (E.coli.) as a core carrier, to develop a novel method for degrading pesticide residues in contaminated soil. We employed a state-of-art light-regulated gene expression system (LightOFF) that can efficiently over-express heterogenous hydrolases that are able to degrade insecticide organophosphorus and fungicide carbendazim respectively, by using darkness as an inducer. This application can lead to two forms of products: enzyme and whole cell products, for efficiently degradation of pesticide residues in soil. In addition, we propose to develop an automatic spraying device to facilitate the usage of whole cell products in real world. Altogether we hope to develop a microbial degradation method with low toxicity, low cost and high efficiency that serves a positive role in maintaining ecological balance.
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<p class="Headline"><span><a href="https://2017.igem.org/Team:SSTi-SZGD/Expression" target="_blank">Expression</a></span></p>
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<p>
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Our LightOFF expression system constitutes a fusion protein of LexA repressor from E. coli SOS regulon, and a blue light sensor (VVD) from Neurospora crassa. Light irradiation causes conformational change of VVD and subsequent dimerization of the fusion protein. The activated dimer thus binds its cognate operator sequence and represses the promoter activity. We first tested its function and induction efficiency.
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<img src="https://static.igem.org/mediawiki/2017/5/55/SSTi-SZGD_Description_Do_Figure.png" alt="Figure"/>
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<p class="Headline"><span><a href="https://2017.igem.org/Team:SSTi-SZGD/Degradation" target="_blank">Degradation</a></span></p>
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<p>
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There are a few identified gene encoding hydrolases to detoxifiy organophosphate and carbendazim pesticides. After a PubMed and iGEM search, we came up with two candidate genes: opd A and mheI. The former has been used in other iGEM projects before and proven working, the latter was conserved in many bacteria species that use carbendazim as a carbon source. We also added a signal peptide from Tat translocase of E. coli to help with protein exportation.
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<p class="subtitle"><a href="https://2017.igem.org/Team:SSTi-SZGD/Degradation" target="_blank">Cell suicide</a></p>
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<p class="subparagraph">
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We considered the risk of releasing GMOs to the natural environment. So we chose to inclulde a suicide gene in the system. Supernova, an engineered genetically-encoded photosensitizer, containing chromphores that generate reactive oxygen species (ROS) upon illumination, and can be used to promote apoptosis in prokaryotic cells. When combine with LightOFF system, darkness induces supernova expression, while light irradiation triggers the release of ROS to promote cell death.
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<p class="subtitle"><a href="https://2017.igem.org/Team:SSTi-SZGD/Applied_Design" target="_blank">Pesticide Residue Sweeper</a></p>
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<p class="subparagraph">
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By combining the above three elements together, we could efficiently produce enzyme (hydrolase) products or construct a live biocatalyst with applications in farms, orchards and gardens. We mainly focused on the product design, target customers, and cost analysis to identify markets. In addition, we developed an automatic spraying device to be used with live biocatalysts.
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<img src="https://static.igem.org/mediawiki/2017/8/8c/SSTi-SZGD_logo_USZ.png" alt="USZ"/>
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<span class="left">A&nbsp;product&nbsp;for&nbsp;the&nbsp;degradation&nbsp;of&nbsp;soil&nbsp;pesticide&nbsp;residues</span>
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<span class="right">Copyright&nbsp;&copy;&nbsp;2017&nbsp;Lucky&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;power&nbsp;by&nbsp;&nbsp;IGEM&nbsp;Team&#58;SSTi-SZGD</span>
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A product&nbsp;for&nbsp;the&nbsp;degradation&nbsp;of&nbsp;soil&nbsp;pesticide&nbsp;residues
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Copyright&nbsp;&copy;&nbsp;2017&nbsp;Lucky&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;power&nbsp;by&nbsp;&nbsp;iGEM&nbsp;Team&#58;SSTi-SZGD
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Latest revision as of 10:13, 1 November 2017

SSTi-SZGD---Description

Pesticide Residue Sweeper--Why We Need it?

The level of pesticide usage in China is 2.5 times above the world average. In June 2016, the total output of chemical pesticides in China was around 3.347 million tons, up by 7.17% compared with the same period of 2015. The deterioration effects of long-term and overdose usage of pesticide in soil include soil erosion, soil contamination, water pollution, organic pollution, reducing biodiversity, etc.
A range of chemical and physical methods have been applied to degrade pesticide residues. The shortfalls of these methods include high in cost, harmful to non-target organisms, food, plants and soils, and likely to cause secondary pollution. We intend to develop a microbial degradation method with low toxicity, low cost and high efficiency to serve a positive role in maintaining ecological balance.

Figure

What we will do?

SSTi-iGEM combined optogenetics and biotechnology, using genetically modified E.coli as a carrier, to improve the microbial method for pesticide residue degradation in soil. We used a light-regulated gene expression system (LightOFF) that efficiently over-express heterogenous hydrolases which degrade insecticide organophosphorus or fungicide carbendazim. This application can result to two forms of products: enzyme and whole cell products. In addition, we propose to develop an automatic praying system that facilitates the application of whole cell products in real-life scenario.

Figure

How to do it?----Project Design

Expression

Our LightOFF expression system constitutes a fusion protein of LexA repressor from E. coli SOS regulon, and a blue light sensor (VVD) from Neurospora crassa. Light irradiation causes conformational change of VVD and subsequent dimerization of the fusion protein. The activated dimer thus binds its cognate operator sequence and represses the promoter activity. We first tested its function and induction efficiency.

Figure

Degradation

There are a few identified gene encoding hydrolases to detoxifiy organophosphate and carbendazim pesticides. After a PubMed and iGEM search, we came up with two candidate genes: opd A and mheI. The former has been used in other iGEM projects before and proven working, the latter was conserved in many bacteria species that use carbendazim as a carbon source. We also added a signal peptide from Tat translocase of E. coli to help with protein exportation.

Cell suicide

We considered the risk of releasing GMOs to the natural environment. So we chose to inclulde a suicide gene in the system. Supernova, an engineered genetically-encoded photosensitizer, containing chromphores that generate reactive oxygen species (ROS) upon illumination, and can be used to promote apoptosis in prokaryotic cells. When combine with LightOFF system, darkness induces supernova expression, while light irradiation triggers the release of ROS to promote cell death.

Pesticide Residue Sweeper

By combining the above three elements together, we could efficiently produce enzyme (hydrolase) products or construct a live biocatalyst with applications in farms, orchards and gardens. We mainly focused on the product design, target customers, and cost analysis to identify markets. In addition, we developed an automatic spraying device to be used with live biocatalysts.

A product for the degradation of soil pesticide residues Copyright © 2017 Lucky      power by  iGEM Team:SSTi-SZGD