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| − | <html lang="en"><head><meta charset="utf-8"/><meta content="width=device-width,initial-scale=1" name="viewport"/><title>Safety | SugarGain | iGEM BITS Goa</title><link href="https://2020.igem.org/Template:BITSPilani-Goa_India/css/contentCSS?action=raw&ctype=text/css" rel="stylesheet"/></head><body><nav id="main-nav"><div class="navbar navbar-expand-lg d-flex justify-content-between bg-none nav-transparent"><a class="navbar-brand d-lg-inline-block" href="https://2020.igem.org/Team:BITSPilani-Goa_India"> Sugar<span>Gain</span></a><div class="controls h-100 d-flex align-items-center"><div class="align-items-center" id="themeSwitchWrapper"><i class="icon-light"></i><div id="themeSwitch"><label class="switch" for="themeSwitchInput"><input id="themeSwitchInput" type="checkbox"/><span class="slider round"></span></label></div><i class="icon-dark"></i></div><button class="btn" id="menuSwitch"><div class="menu-icon"></div></button></div></div><div id="fullscreenMenuWrapper"><div 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| | }</style><header class="d-flex justify-content-center"><div class="container d-flex align-items-end justify-content-between"><div id="heading"><h1>Safety</h1></div><a href="https://unsplash.com/photos/lZ4xZZuk8iA" rel="noopener" target="_blank"><span data-placement="top" id="bg-attribution" title="Photo by Josh Withers on Unsplash"></span></a></div></header><main class="hello theme-yellow"><div class="container"><div class="row"><div class="sidebar"><div class="nav" id="contents"><ul></ul></div></div><div class="content"><article><p>Biosafety is the ethical obligation of a bioengineer. Throughout our project design we have tried to be as mindful as possible regarding identifying strategies for robust biosafety. Since our project is closely related to a consumable sugar, we have devised a three-tier fail-safe mechanism after taking into consideration the various use cases of our chassis.</p><h1>Level 1: Kill Switch</h1><p>We intend to introduce our strain of bacteria into the sugarcane matrix to synthesise anti-invertase. Considering the close proximity of the system to both agricultural environments and human consumables, it is indispensable that our bacteria include a lethality mechanism. We propose to use a modified type II ccdA-ccdB toxin-antitoxin system as our kill switch that is activated upon exposure to atmospheric concentrations of oxygen. A change in the oxygen concentration in the environment results in a sharp increase in the amount of the toxin protein being synthesised which disturbs the already established equilibrium between the toxin and antitoxin, effectively killing the cell, and thus preventing any contamination by stray microbes. More information about the design of the kill switch can be found <a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Design#KillSwitch">here</a> and a study of its dynamics <a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Model/Kill_Switch">here</a>.</p><h1>Level 2: Auxotropic Regulation</h1><p>There are several amino acids that have been detected in sugarcane including, pipecolic acid and hydroxypipecolic acid. Increasing the supply of nitrogen increases markedly the amides asparagine and glutamine and less markedly the basic amino‐acids lysine, histidine, arginine and tryptophan. The neutral and acidic amino‐acids tend to increase and then decrease in level with increasing nitrogen supply. An auxotroph of tryptophan would bleh. You can find a detailed description of auxotrophic model <a href="">here</a></p><h1>Level 3: Sugar Manufacturing Process</h1><blockquote><p>Need better quality image of the industy process</p></blockquote><p>During the sugar manufacturing process the cane juice is heated at the evaporation station to temperatures greater than 100°C for upto eught hours. Furthermore, sulphur dioxide is passed through the solution at the clarifying stage along with a change in pH. We have designed experiments to mimic these industrial conditions on a lab scale. More details about the same can be found <a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Experiments">here</a>. The bacteria would not be able to handle the stress of these conditions and hence the manufacturing process would act as a third failsafe.</p></article><div class="d-flex justify-content-between my-5"><a class="button prev flex justify-content-end long" href="https://2020.igem.org/Team:BITSPilani-Goa_India/Engineering"><span aria-hidden="" class="circle"><span class="icon arrow"></span></span><span class="button-text">Engineering</span></a><a class="button next super-long" href="https://2020.igem.org/Team:BITSPilani-Goa_India/Implementation"><span aria-hidden="" class="circle"><span class="icon arrow"></span></span><span class="button-text">Implementation</span></a></div></div></div></div></main><footer><div id="footerNav"><div class="container"><div class="row justify-content-between"><div class="col-sm-6 col-md-4 col-lg-3 py-2"><a class="footer-heading">Project</a><ul><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Description">Description</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Design">Design</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Model">Model</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Engineering">Engineering</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Safety">Safety</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Implementation">Implementation</a></li></ul></div><div class="col-sm-6 col-md-4 col-lg-3 py-2"><a class="footer-heading">Human Practices</a><ul><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Human_Practices">Human-Centered Design</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Education">Science Communication</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Humans_of_iGEM">Humans of iGEM</a></li><li class="py-1"><a 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class="footer-heading">Team</a><ul><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Team">Team</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Attributions">Attributions</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Collaborations">Collaborations</a></li></ul></div><div class="col-sm-6 col-md-4 col-lg-3 py-2"><a class="footer-heading">Contribution</a><ul><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Contribution">Overview</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Software/Wiki">Wiki Software</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Documentation">Documentation</a></li></ul></div><div class="col-sm-6 col-md-4 col-lg-3 py-2"><a class="footer-heading">Judging</a><ul><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Judging/Bronze">Bronze</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Judging/Silver">Silver</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Judging/Gold">Gold</a></li></ul></div><div class="col-sm-6 col-md-4 col-lg-3 py-2"><a class="footer-heading">Other Areas</a><ul><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Excellence">Overview</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Dance">Dance</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Policy">Policy</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Entrepreneurship">Entrepreneurship</a></li></ul></div></div></div></div><div id="footerBrand"><div class="container"><div class="row"><div id="footerLogo"></div></div><div class="row justify-content-center" id="footerIcons"><a class="gmail" href="mailto:igembitsgoa@gmail.com"></a><a class="insta" href="https://instagram.com/igem_bits" rel="noopener" target="_blank"></a><a class="twitter" href="https://twitter.com/igembitsgoa" rel="noopener" target="_blank"></a></div></div></div></footer><script src="https://2020.igem.org/Template:BITSPilani-Goa_India/content-bundleJS?action=raw&ctype=text/javascript"></script></body></html> | | }</style><header class="d-flex justify-content-center"><div class="container d-flex align-items-end justify-content-between"><div id="heading"><h1>Safety</h1></div><a href="https://unsplash.com/photos/lZ4xZZuk8iA" rel="noopener" target="_blank"><span data-placement="top" id="bg-attribution" title="Photo by Josh Withers on Unsplash"></span></a></div></header><main class="hello theme-yellow"><div class="container"><div class="row"><div class="sidebar"><div class="nav" id="contents"><ul></ul></div></div><div class="content"><article><p>Biosafety is the ethical obligation of a bioengineer. Throughout our project design we have tried to be as mindful as possible regarding identifying strategies for robust biosafety. Since our project is closely related to a consumable sugar, we have devised a three-tier fail-safe mechanism after taking into consideration the various use cases of our chassis.</p><h1>Level 1: Kill Switch</h1><p>We intend to introduce our strain of bacteria into the sugarcane matrix to synthesise anti-invertase. Considering the close proximity of the system to both agricultural environments and human consumables, it is indispensable that our bacteria include a lethality mechanism. We propose to use a modified type II ccdA-ccdB toxin-antitoxin system as our kill switch that is activated upon exposure to atmospheric concentrations of oxygen. A change in the oxygen concentration in the environment results in a sharp increase in the amount of the toxin protein being synthesised which disturbs the already established equilibrium between the toxin and antitoxin, effectively killing the cell, and thus preventing any contamination by stray microbes. More information about the design of the kill switch can be found <a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Design#KillSwitch">here</a> and a study of its dynamics <a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Model/Kill_Switch">here</a>.</p><h1>Level 2: Auxotropic Regulation</h1><p>There are several amino acids that have been detected in sugarcane including, pipecolic acid and hydroxypipecolic acid. Increasing the supply of nitrogen increases markedly the amides asparagine and glutamine and less markedly the basic amino‐acids lysine, histidine, arginine and tryptophan. The neutral and acidic amino‐acids tend to increase and then decrease in level with increasing nitrogen supply. An auxotroph of tryptophan would bleh. You can find a detailed description of auxotrophic model <a href="">here</a></p><h1>Level 3: Sugar Manufacturing Process</h1><blockquote><p>Need better quality image of the industy process</p></blockquote><p>During the sugar manufacturing process the cane juice is heated at the evaporation station to temperatures greater than 100°C for upto eught hours. Furthermore, sulphur dioxide is passed through the solution at the clarifying stage along with a change in pH. We have designed experiments to mimic these industrial conditions on a lab scale. More details about the same can be found <a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Experiments">here</a>. The bacteria would not be able to handle the stress of these conditions and hence the manufacturing process would act as a third failsafe.</p></article><div class="d-flex justify-content-between my-5"><a class="button prev flex justify-content-end long" href="https://2020.igem.org/Team:BITSPilani-Goa_India/Engineering"><span aria-hidden="" class="circle"><span class="icon arrow"></span></span><span class="button-text">Engineering</span></a><a class="button next super-long" href="https://2020.igem.org/Team:BITSPilani-Goa_India/Implementation"><span aria-hidden="" class="circle"><span class="icon arrow"></span></span><span class="button-text">Implementation</span></a></div></div></div></div></main><footer><div id="footerNav"><div class="container"><div class="row justify-content-between"><div class="col-sm-6 col-md-4 col-lg-3 py-2"><a class="footer-heading">Project</a><ul><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Description">Description</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Design">Design</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Model">Model</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Engineering">Engineering</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Safety">Safety</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Implementation">Implementation</a></li></ul></div><div class="col-sm-6 col-md-4 col-lg-3 py-2"><a class="footer-heading">Human Practices</a><ul><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Human_Practices">Human-Centered Design</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Education">Science Communication</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Humans_of_iGEM">Humans of iGEM</a></li><li class="py-1"><a 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class="footer-heading">Team</a><ul><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Team">Team</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Attributions">Attributions</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Collaborations">Collaborations</a></li></ul></div><div class="col-sm-6 col-md-4 col-lg-3 py-2"><a class="footer-heading">Contribution</a><ul><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Contribution">Overview</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Software/Wiki">Wiki Software</a></li><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Documentation">Documentation</a></li></ul></div><div class="col-sm-6 col-md-4 col-lg-3 py-2"><a class="footer-heading">Judging</a><ul><li class="py-1"><a href="https://2020.igem.org/Team:BITSPilani-Goa_India/Judging/Bronze">Bronze</a></li><li class="py-1"><a 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