Difference between revisions of "Team:REC-CHENNAI/Description"

 
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<h1>ANTIMICROBIAL PEPTIDES- A SOLUTION</h1>
 
<h1>ANTIMICROBIAL PEPTIDES- A SOLUTION</h1>
 
<p>AMPs are gaining popularity due to their effectiveness against a wide range of pathogenic organisms that are resistant to conventional drugs. Since their mode of action, for the most part, exploits general but fundamental structural characteristics such as the bacterial cell membrane and in many cases they may have multiple targets within cells, the likelihood of the emergence of resistance is thought to be considerably reduced compared with that for many current antibiotics, which have more specific molecular targets (Håvard Jenssen et al).</p>
 
<p>AMPs are gaining popularity due to their effectiveness against a wide range of pathogenic organisms that are resistant to conventional drugs. Since their mode of action, for the most part, exploits general but fundamental structural characteristics such as the bacterial cell membrane and in many cases they may have multiple targets within cells, the likelihood of the emergence of resistance is thought to be considerably reduced compared with that for many current antibiotics, which have more specific molecular targets (Håvard Jenssen et al).</p>
<p>For easy biosynthetic production, we decided to work with a small peptide, devoid of cysteine knots. In addition, we preferred that it not be easily degraded by proteases. Keeping these in mind, we looked for peptides that possess antimicrobial activity .Of the many that we came across, we were interested in working Latarcin2a.</p>
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<p>For easy biosynthetic production, we decided to work with a small peptide, devoid of cysteine knots. In addition, we preferred that it not be easily degraded by proteases. Keeping these in mind, we looked for peptides that possess antimicrobial activity .Of the many that we came across, we were interested in working Lt2a.</p>
 
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<img src="https://static.igem.org/mediawiki/2017/2/25/Rec_chennai_description3.jpg">
 
<img src="https://static.igem.org/mediawiki/2017/2/25/Rec_chennai_description3.jpg">
<p>position of the F10 in the peptide is shown in green color.</p>
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<p>Position of the F10 in the peptide is shown in green color.</p>
 
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                       <p> At low cell density, the quantity of AHL produced is too little for it to bring about Latarcin production. As the cell density increases, the cells divide and simultaneously produce both AHL and LuxR. Without the AHL, LuxR is unstable and gets degraded shortly after production. But beyond a certain threshold level, AHL begins forming a complex with LuxR, preventing its degradation. This complex now binds to the Lux promoter that regulates Latarcin synthesis. On binding, the promoter immediately switches ON and begins to produce a large amount of Latarcin. A small portion of the synthesised Latarcin destroys the host, and the rest can now be isolated and purified. </p>
 
                       <p> At low cell density, the quantity of AHL produced is too little for it to bring about Latarcin production. As the cell density increases, the cells divide and simultaneously produce both AHL and LuxR. Without the AHL, LuxR is unstable and gets degraded shortly after production. But beyond a certain threshold level, AHL begins forming a complex with LuxR, preventing its degradation. This complex now binds to the Lux promoter that regulates Latarcin synthesis. On binding, the promoter immediately switches ON and begins to produce a large amount of Latarcin. A small portion of the synthesised Latarcin destroys the host, and the rest can now be isolated and purified. </p>
  
<img src="https://static.igem.org/mediawiki/2017/a/a6/Igem_recchennai_2017_quorom.png"/>
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<center><img src="https://static.igem.org/mediawiki/2017/a/a6/Igem_recchennai_2017_quorom.png"/></center>
 
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<h1>APPLICATION</h1>
 
<h1>APPLICATION</h1>
<p>Text here</p>
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<p> It is estimated that food worth $1 trillion is wasted every year. This accounts for about one third of the world's food. Of this loss, 21% is caused due to microbial food spoilage. Thus finding a niche in the food preservation sector, we decided to develop a food wrapper that can extend the shelf life of mainly fruits and vegetables. </p>
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  <p>  Inspired by 2012 iGEM Team Groningen, we propose a delivery system containing spores of genetically engineered <i>Bacillus</i>  enclosed within two layers of PVDF membrane. (We chose to work with <i> E.coli </i> to carry out cloning experiments , due to its ease of handling. However, we propose that <i> Bacillus</i> spores be used in making the Food wrapper) The system is designed in such a way that only the PVDF membrane in contact with food will allow Latarcin to pass through. The purpose of the outer membrane(filter) is to prevent the entry of microorganisms, thus avoiding chances of further contamination. </p>
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  <p>  To serve as a nutrient source for the bacteria, LB broth is contained within a compartment made of PVC, which is sandwiched between the two PVDF membranes. When mechanical stress is applied, the delicate PVC membrane disintegrates, allowing the bacteria to come in contact with the LB broth. When this happens, the dormant bacteria start multiplying.  Once the bacteria reach a substantial number,  due to the quorum sensing mechanism, the mutant latarcin will be  synthesised. </p>
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<p>  This results in abundant production of Latarcin by the host bacteria.  Latarcin diffuses through the inner PVDF membrane and reaches the food. On encountering bacteria present on the food, Latarcin effectively kills them via the carpet mechanism. Because of its half life of 10 hours, any unused Latarcin is therefore degraded, making the food safe for human consumption. </p>
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<center><img src="https://static.igem.org/mediawiki/2017/1/17/Igem_recchennai_2017_projectapp.png" style="width:50%"/></centeR>
 
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Latest revision as of 20:20, 1 November 2017