Difference between revisions of "Team:Kent"

 
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<a style="opacity:0.1"; width="200px" class="twitter-timeline" href="https://twitter.com/igem_kent" data-widget-id="615512808541786112" data-chrome="transparent">Tweets by @igem_kent</a>
 
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  <h1 align="center"> Welcome to our iGem page </h1>
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<a name="#ref"></a> <h1 align="center"> Developing Green Nanowire </h1>
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Our project aims to engineer a novel synthetic biology solution to the production of conductive nano-wires. Our system takes advantage of the endogenous curli system of E.coli to produce functional extracellular amyloid nano-fibrils composed of the amyloid forming domain of the yeast prion protein Sup35NM. The curli system uses the Sec transport pathway to translocate Sup35NM tagged with the Curli-signal sequence through the inner membrane where it is transported from the periplasm through a specific pore to the outer membrane where it assembles into repeating units. We intend to engineer a protein containing Sup35NM (containing the amyloid forming domain of Sup35) linked together to cytochrome <i>b</i><sub>562</sub> that will transport electrons along the amyloid fibrils. The folding of cytochrome <i>b</i><sub>562</sub> requires the cofactor haem, which we are able to add to the growth medium to facilitate folding extra cellular. The wider aim of the project is to funnel electrons from the electron transport chain into the sup35NM/cytochrome <i>b</i><sub>562</sub> nanowire, which together would provide a renewable source of energy that could be used to power small consumer products.</p>
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With rapid technological advancement in the energy, electrical and computing industries there is pressure to increase processing power while downscaling circuitry, and at the same time make components and circuitry biocompatible for medical or biological applications. The recent advances in fabrication of nano-wires have fuelled the need for biocompatible wires that can interface with cellular components. Currently most electronic components use copper clad laminates, and rare earth metals, which are finite resources and require significant amount of space, energy, metals and rare resources. Nano-wires formed from proteins by bacteria provide a solution to the fabrication of a Nano-material in terms of miniaturization, improved efficiency, renewable use of energy and materials, and biocompatibilty.
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<h1 align="center">How we’re going to do it</h1>
 
  
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<p align="center">We are going to utilise the endogenous Curli system of E.coli VS45 strain to export a protein that will form amyloid nano-fibres. The protein will be expressed from a plasmid that we are going to engineer, containing sequence coding for the Sup35NM protein and cytochrome <i>b</i><sub>562</sub> gene inserted into the expression site. The curli system consists of csgG export system, which recognises the csgA N-terminal signal sequence that is attached to the sup35NM protein. This will allow the sup35NM/cytochrome <i>b</i><sub>562</sub> chimera to be translocated to the extracellular space to form an amyloid fibre. 

To reach the curli-specific pore on the outer membrane the sup35NM/cytochrome <i>b</i><sub>562</sub> complex must pass the inner membrane to the periplasm via the Sec translocation pathway. Cytochrome <i>b</i><sub>562</sub> requires the cofactor haem to allow correct folding. Cofactors cannot bind before translocation via the Sec pathway, however cytochrome <i>b</i><sub>562</sub> can bind to haem that is exogenously added to the solution after export, thus allowing the cytochrome to fold into its active conformation extracellularly. The cytochrome on the amyloid fibres will then transport electrons down the transport chain, acting as a nanowire. 

We will first create a biofilm of conducting nano-fibres over a surface that we would like electricity to flow across. This will result in a conducting material connecting cells that could be used as a replacement for conventional nanowire allowing the downscaling of consumer products. The ultimate goal of the project is to funnel electrons from the electron transport chain of the bacterial respiratory chain into the amyloid chain. This would allow the production of a self-powering unit that aggregates its own nanowire.</p>
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</a><h1 align="center">Our Answer To Biocompatible Nanowire</h1>
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Our amyloid nano-wire provides an alternative material and method of producing nano-wire, whilst being environmentally friendly as it is a renewable resource and can self-assemble. We utilized the amyloid forming protein, Sup35-NM, which can be exported by <i>E. coli</i> into a solution, where the protein self-assembles to form amyloid fibrils. Amyloid fibrils are well-suited for use as nanowire due to their high heat stability, mechanical properties, biocompatibility, and ease of functionalization.</p>
 
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<h1 align="center">Future</h1>
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With further engineering, in the future it would be possible to insert an electron carrier into the periplasm, allowing electrons from the electron transport chain to be sequestered into the amyloid nanowire directly. This could be achieved using strains of <i>E.coli</i> that allow amyloid fibres to bind to the outside of the cell at specific points. Ultimately this will allow self-powering bio-electronic devices that could be used in products such as mobile phones, energy plants that generates green bio-energy, and small self-contained batteries that generate and transport its own electricity and eliminates the need for chargers. </p>
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Latest revision as of 21:25, 18 September 2015


iGEM Kent 2015





























Developing Green Nanowire

With rapid technological advancement in the energy, electrical and computing industries there is pressure to increase processing power while downscaling circuitry, and at the same time make components and circuitry biocompatible for medical or biological applications. The recent advances in fabrication of nano-wires have fuelled the need for biocompatible wires that can interface with cellular components. Currently most electronic components use copper clad laminates, and rare earth metals, which are finite resources and require significant amount of space, energy, metals and rare resources. Nano-wires formed from proteins by bacteria provide a solution to the fabrication of a Nano-material in terms of miniaturization, improved efficiency, renewable use of energy and materials, and biocompatibilty.

































Our Answer To Biocompatible Nanowire

Our amyloid nano-wire provides an alternative material and method of producing nano-wire, whilst being environmentally friendly as it is a renewable resource and can self-assemble. We utilized the amyloid forming protein, Sup35-NM, which can be exported by E. coli into a solution, where the protein self-assembles to form amyloid fibrils. Amyloid fibrils are well-suited for use as nanowire due to their high heat stability, mechanical properties, biocompatibility, and ease of functionalization.






























Future

With further engineering, in the future it would be possible to insert an electron carrier into the periplasm, allowing electrons from the electron transport chain to be sequestered into the amyloid nanowire directly. This could be achieved using strains of E.coli that allow amyloid fibres to bind to the outside of the cell at specific points. Ultimately this will allow self-powering bio-electronic devices that could be used in products such as mobile phones, energy plants that generates green bio-energy, and small self-contained batteries that generate and transport its own electricity and eliminates the need for chargers.