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<h1><font color="#AC2B37">Project Summary</font></h1>
 
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<h3>Two Bacteria = Two Solutions</h3>
 
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<p>Our project seeks to engineer two separate strains of bacterium to solve problems related to lead pollution. Our first bacterium, <i>Bacillus subtilis</i>, will function as a biosensor by changing colors to indicate how much lead contamination, if any, is present in a sample of water. If lead is present, our second bacterium, <i>Lactobacillus rhamnosus GG</i>, can be taken daily as a probiotic to potentially reduce the amount of lead contamination in ingested water. </p>
 
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<h3>Got Lead?</h3>
 
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<p>Lead contamination in drinking water is becoming a larger problem due to the aging infrastructure present in America and other countries with lead pipes (Ciccatelli). Our project was namely inspired by the water crisis in Flint, Michigan, which has left many of the city’s inhabitants without drinkable tap water since 2014 (Flint). While the problem in Flint certainly is extreme, lead pollution is actually found all over the US, including Massachusetts (Globe). If there is a lead pipe anywhere between the source of your water and your faucet, you could have lead contamination. <br/> </p>
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<p>Lead contamination in drinking water is a major problem across the U.S. Due to aging infrastructure, many people lack access to a consistent, clean source of water in their homes. Cities and local governments must react urgently to lead contamination and administer water testing and treatment solutions to their citizens. Our project aims to improve lead testing and treatment in emergency situations by developing a lead biosensor and probiotic, as well as a lead assay. </p>
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<p>Unfortunately, many lead detection products today only offer a ‘yes/no’ detection system, set off if the quantity of lead in the sample is above a certain level. Even once lead is detected, homeowners and renters alike may be unable to change the source of their water due to financial and other restrictions associated with lead filtration and other methods available today. Our project aims to overcome these problems, while affordably and accessibly providing answers to the questions “Is there lead in my water, and how much?” as well as “What can I do about the lead right now?” </p>
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<h3>Two Bacteria, Two Solutions</h3>
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<p>Our design of the at-home lead biosensor considers the safety of the organism as well as a quantitative yet straightforward indication of lead contamination. The biosensor colorimetrically signals benchmark levels of lead contamination in a sample of water by producing specific chromoproteins. These benchmarks are based on EPA standards for lead contamination in water and confirmed by our lead assay. For our biosensor to express chromoproteins at these benchmarks, we utilized error-prone PCR to mutate an existing lead promoter; specific promoters were tested and selected based on their sensitivity to lead. We are conducting proof-of-principle demonstrations of the biosensor in <i>Escherichia coli</i>, and will ultimately transform it into the Generally Recognized As Safe organism <i>Bacillus subtilis</i>. <br/> </p>
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<p>Our probiotic, <i>Lactobacillus rhamnosus</i>, offers an emergency prophylactic treatment for lead contamination by absorbing lead from the gastrointestinal tract. Someone who must continue to consume contaminated water would simply eliminate the leaded bacteria with the rest of their waste. To achieve this, we are using selective pressure to evolve a probiotic with enhanced lead-binding capacity. The extent to which the probiotic absorbed lead was analyzed by our lead assay. Our project will improve the accessibility of lead detection and bioremediation for the general population. </p>
  
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<h3>The Engineering Behind it All</h3>
 
<h3>The Engineering Behind it All</h3>
 
<h5><font color="#A9B0B7">Bio-Detection</font></h5>
 
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<p>Our <i>Bacillus subtilis</i> biosensor will contain a plasmid with three chromoprotein genes, each with a slightly different version of a promoter which is responsive to lead concentration. This will result in three different colors to be produced at different concentrations of lead: low, medium, and high. This will help users understand that they have lead in their water in addition to how much. <br/> </p>
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Our <i>Bacillus subtilis</i> biosensor will contain a plasmid with three chromoprotein genes, each with a slightly different version of a promoter which is responsive to lead concentration. This will result in three different colors to be produced at different concentrations of lead: low, medium, and high. This will help users understand that they have lead in their water in addition to how much.  
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<img src="https://static.igem.org/mediawiki/2017/7/73/T--WPI_Worcester--2017-Biosensor_Beaker.png" alt="Biosensor Beaker" style="float:left;width:800px;height:415px;">
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<h5><font color="#A9B0B7">Probiotic Purification</font></h5>
 
<h5><font color="#A9B0B7">Probiotic Purification</font></h5>
 
<p><i>Lactobacillus rhamnosus</i> has natural lead-binding capabilities (Marc). When <i>Lactobacillus</i> binds lead, it keeps the lead from being absorbed by your system, allowing the normally toxic metal to pass harmlessly out of your body by just taking one pill every day. We plan to “force-evolve” <i>Lactobacillus rhamnosus GG</i> to have enhanced lead-binding capabilities as well as the ability to survive in higher concentrations of lead by exposing it to the metal and observing its growth.<br/><br/></p>
 
<p><i>Lactobacillus rhamnosus</i> has natural lead-binding capabilities (Marc). When <i>Lactobacillus</i> binds lead, it keeps the lead from being absorbed by your system, allowing the normally toxic metal to pass harmlessly out of your body by just taking one pill every day. We plan to “force-evolve” <i>Lactobacillus rhamnosus GG</i> to have enhanced lead-binding capabilities as well as the ability to survive in higher concentrations of lead by exposing it to the metal and observing its growth.<br/><br/></p>
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<h5> Sources:</h5>
 
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<li>Ciccatelli, Amanda. “How America’s Aging Infrastructure is Creating More Toxic Lawsuits.” Inside Counsel. ALM Media, 30 March 2017. Web. 23 June 2017. </li>
 
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<li>"Flint Water Crisis Fast Facts." CNN. Cable News Network, 10 Apr. 2017. Web. 16 June 2017. </li>
 
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<li>Globe, The Boston. "High lead levels found in water at 160 Mass. schools." Boston.com. The Boston Globe, 16 Nov. 2016. Web. 16 June 2017. </li>
 
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<li>Marc, and Jeremy P. Burtona And. "Possible Role of Probiotics" Applied and Environmental Microbiology. N.p., 01 Sept. 2012. Web. 16 June 2017. </li>
 
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Latest revision as of 20:08, 13 October 2017




Project Summary


Got Lead?

Lead contamination in drinking water is a major problem across the U.S. Due to aging infrastructure, many people lack access to a consistent, clean source of water in their homes. Cities and local governments must react urgently to lead contamination and administer water testing and treatment solutions to their citizens. Our project aims to improve lead testing and treatment in emergency situations by developing a lead biosensor and probiotic, as well as a lead assay.

Two Bacteria, Two Solutions

Our design of the at-home lead biosensor considers the safety of the organism as well as a quantitative yet straightforward indication of lead contamination. The biosensor colorimetrically signals benchmark levels of lead contamination in a sample of water by producing specific chromoproteins. These benchmarks are based on EPA standards for lead contamination in water and confirmed by our lead assay. For our biosensor to express chromoproteins at these benchmarks, we utilized error-prone PCR to mutate an existing lead promoter; specific promoters were tested and selected based on their sensitivity to lead. We are conducting proof-of-principle demonstrations of the biosensor in Escherichia coli, and will ultimately transform it into the Generally Recognized As Safe organism Bacillus subtilis.

Our probiotic, Lactobacillus rhamnosus, offers an emergency prophylactic treatment for lead contamination by absorbing lead from the gastrointestinal tract. Someone who must continue to consume contaminated water would simply eliminate the leaded bacteria with the rest of their waste. To achieve this, we are using selective pressure to evolve a probiotic with enhanced lead-binding capacity. The extent to which the probiotic absorbed lead was analyzed by our lead assay. Our project will improve the accessibility of lead detection and bioremediation for the general population.

The Engineering Behind it All

Bio-Detection

Our Bacillus subtilis biosensor will contain a plasmid with three chromoprotein genes, each with a slightly different version of a promoter which is responsive to lead concentration. This will result in three different colors to be produced at different concentrations of lead: low, medium, and high. This will help users understand that they have lead in their water in addition to how much. Biosensor Beaker


















Probiotic Purification

Lactobacillus rhamnosus has natural lead-binding capabilities (Marc). When Lactobacillus binds lead, it keeps the lead from being absorbed by your system, allowing the normally toxic metal to pass harmlessly out of your body by just taking one pill every day. We plan to “force-evolve” Lactobacillus rhamnosus GG to have enhanced lead-binding capabilities as well as the ability to survive in higher concentrations of lead by exposing it to the metal and observing its growth.

Probiotic Beaker