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<h2>UNITN iGEM 2015</h2>
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<p>Electron-production with <br /> engineered <i>E. coli</i> and <br /> <strong>proteorhodopsin</strong></p>
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<h2><strong>Work hard, Work safe</strong></h2>
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<h2>Electron production with <strong><i>E. coli</i></strong> and proteorhodopsin </h2>
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<p>The aim of our project is to develop a pseudoautotrophic <span class="italicEnph">E. coli</span>, enabling it to live in the microbial fuel cell anaerobic condition. Our idea is to engineer E. coli with proteorhodopsin, a light-powered proton pump identified in bacteria that live in deep waters. This protein is activated by light and needs retinal molecules to pump protons outward following excitation. Thus we want to engineer our bacteria with blh, that encodes for the enzyme that cleaves beta-carotene and produces retinal.</p>
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<p>To increase electron production we plan to overexpress pncB that represents the limiting step in the NAD+ synthesis pathway. The more NAD+ is produced, the more electrons are transported by NADH. In short: light activates proteorhodopsin, increases bacterial viability, and drives to a more efficient electron production. Our bacterial device will give rise to a self-sustainable system, powered by sunlight during the day and by a light bulb at night. Daytime-electricity can be either used or stored to be employed as a power source for the lightbulb at night.</p>
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<p class="pintest">Electricity from E. coli<br/>
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<span>Check out our amazing idea! Have fun!<br/>
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<p class="pintest">Electricity from E. coli<br/>
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<span>Check out our amazing idea! Have fun!<br/>
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<a class="btn" href="#">Project</a></span></p>
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<li style="width: 25%; background-image: url('https://static.igem.org/mediawiki/2015/f/fc/Unitn_pics_slider_3.jpg'); ">
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<p class="pintest">Electricity from E. coli<br/>
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<span>Check out our amazing idea! Have fun!<br/>
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<a class="btn" href="#">Project</a></span></p>
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<span>We are UNITN iGEM 2015 and we are such essence as dNTPs are made on<br/>
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<span>Many techniques, different strains and so much fluorescence… Enjoy the Measurement Interlab Study<br/>
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<a class="btn" href="#"> To->Interlab </a></span></p>
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<p class="pintest">Synthetic Biology and mountains<br/>
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<p>We have reviewed our organisms and parts against the <a class="i_linker" href="https://2015.igem.org/Safety/WhiteList" target="_blank">White List</a>. All the parts and bacterial strains that we used are in the <span class="i_enph">Risk Group 1</span></p>
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<p>We have submitted to iGEM headquarters <span class="i_enph">“About Our Lab”</span> and <span class="i_enph">“About Our Project”</span> questionnaires by the June 26<sup>th</sup> deadline</p>
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<p>We are participating also to the Measurements Interlab Study. Check out our progress in the Interlab Section!</p>
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<p>We have submitted to iGEM headquarters the <span class="i_enph">“Final Safety Form”</span> by the August 28<sup>th</sup> deadline</p>
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<p>All the parts we used throughout the project are NOT toxic or dangerous for  humans or the environment. We neither worked with proteins toxic themselves, nor we used enzymes that synthesize toxic molecules. In addition to basic parts from the registry (i.e. RBS, promoters, β-carotene, etc), we also used two genes from the SAR86 uncultured bacteria. These genes are proteorhodopsin, which was extracted from the Registry (BBa_K773002) and blh (15,15’-β-carotene dioxygenase), which was synthesized by Genescript. There is no associated risk level with the uncultured bacteria SAR86, however we are using only DNA sequences that encodes for genes, which carry distinct functions that are no harmful to humans, or the environment. We have clarified this issue with iGEM Safety Office.</p>
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<p>All the bacterial strains used are from Risk Group 1. We used the following strains:</p>
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<p>Last but non least we adopted the appropriate PPE during the lab work.  We always used lab coats, nitrile gloves and goggles.  We safely operated with biological material under the laminar flow cabinet BioAir SAFEMATE Series Class II 1.8.</p>
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<p>We used chemicals that are associated with hazard (ethidium bromide, azide,  organic solvents) under the chemical hood Fume Hood MOD. ASEM® 120EN New - Class 0.</p>
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Revision as of 12:14, 10 August 2015

Electron production with E. coli and proteorhodopsin


The aim of our project is to develop a pseudoautotrophic E. coli, enabling it to live in the microbial fuel cell anaerobic condition. Our idea is to engineer E. coli with proteorhodopsin, a light-powered proton pump identified in bacteria that live in deep waters. This protein is activated by light and needs retinal molecules to pump protons outward following excitation. Thus we want to engineer our bacteria with blh, that encodes for the enzyme that cleaves beta-carotene and produces retinal.

To increase electron production we plan to overexpress pncB that represents the limiting step in the NAD+ synthesis pathway. The more NAD+ is produced, the more electrons are transported by NADH. In short: light activates proteorhodopsin, increases bacterial viability, and drives to a more efficient electron production. Our bacterial device will give rise to a self-sustainable system, powered by sunlight during the day and by a light bulb at night. Daytime-electricity can be either used or stored to be employed as a power source for the lightbulb at night.

Achievements

# new BioBricks

We built and submitted a total number of new BioBricks and characterized # of them. Check out our [Parts] page to know more!

Improved # Parts

Something existing, reshaped better. # now works!

Measurements Interlab Study

We are participating also to the Measurements Interlab Study. Check out our progress in the Interlab Section!