Difference between revisions of "Team:ShanghaitechChina/Demonstrate"
m |
m |
||
| Line 15: | Line 15: | ||
<div class="row"> | <div class="row"> | ||
<div class="col-lg-12"> | <div class="col-lg-12"> | ||
| − | <h1 align="center">Demonstration of Solar Hunter in a Nutshell</h1> | + | <h1 align="center" id="Demonstration ">Demonstration of Solar Hunter in a Nutshell</h1> |
<p></p> | <p></p> | ||
Artificial photosynthesis represents a promising solution for energy issues, however, the efficiency, robustness, and scalability does not meet the requirements of industrial applications. We proposed and demonstrated a sun-powered biofilm-interfaced artificial hydrogen-producing system, Solar Hunter, that could potentially solve the issues above. Biofilm-anchored nanorods can efficiently convert photons to electrons, which seamlessly tap into the electron chain of engineered strain carrying FeFe hydrogenase gene cluster, thereby achieving high-efficiency hydrogen production. Furthermore, the intrinsic adherence of biofilms towards various interfaces allows us to grow biofilms on easy-separation micro-beads, therefore facilitating recyclable usage of the biofilm-anchored NRs and endowing this whole system with recyclability. Notably, our hydrogen production has shown great stability compared to some precursors using hydrogenase. Such system can also be adapted to other energy-oriented applications by utilizing engineered new strains with a diverse spectrum of enzymes or metabolic pathways. In this section, we show the successful hydrogen production data with all the components. For the thorough tour of hydrogen production assays we did, please refer to Integrative Bio-hydrogen System (https://2016.igem.org/Team:ShanghaitechChina/IBS). <p></p> | Artificial photosynthesis represents a promising solution for energy issues, however, the efficiency, robustness, and scalability does not meet the requirements of industrial applications. We proposed and demonstrated a sun-powered biofilm-interfaced artificial hydrogen-producing system, Solar Hunter, that could potentially solve the issues above. Biofilm-anchored nanorods can efficiently convert photons to electrons, which seamlessly tap into the electron chain of engineered strain carrying FeFe hydrogenase gene cluster, thereby achieving high-efficiency hydrogen production. Furthermore, the intrinsic adherence of biofilms towards various interfaces allows us to grow biofilms on easy-separation micro-beads, therefore facilitating recyclable usage of the biofilm-anchored NRs and endowing this whole system with recyclability. Notably, our hydrogen production has shown great stability compared to some precursors using hydrogenase. Such system can also be adapted to other energy-oriented applications by utilizing engineered new strains with a diverse spectrum of enzymes or metabolic pathways. In this section, we show the successful hydrogen production data with all the components. For the thorough tour of hydrogen production assays we did, please refer to Integrative Bio-hydrogen System (https://2016.igem.org/Team:ShanghaitechChina/IBS). <p></p> | ||
| Line 29: | Line 29: | ||
<ul id="sidebar" class="nav bs-docs-sidenav "> | <ul id="sidebar" class="nav bs-docs-sidenav "> | ||
<li > | <li > | ||
| − | <a href="# | + | <a href="#Demonstration ">Demonstration of Solar Hunter in a Nutshell</a> |
</li> | </li> | ||
<li> | <li> | ||
| − | <a href="# | + | <a href="#Method" style="font-size:14px;margin-left:15px;">Method and Instrument</a> |
| − | + | ||
| − | + | ||
| − | + | ||
</li> | </li> | ||
<li> | <li> | ||
| Line 48: | Line 45: | ||
<div class="row"> | <div class="row"> | ||
<div class="col-lg-12"> | <div class="col-lg-12"> | ||
| − | + | <center> <h1 id="Method"> Method and Instrument</h1></center> | |
| + | <h2> Method </h2> | ||
| + | The biofilm, whose subunit was CsgA engineered with HisTag on N-termial and SpyCachter-HisTag on C-terminal, was grown on microspheres, 25 micrometers in diameter for 48 hours. NR’s (7.72*10^-9 M) were then added and given 30 min to bind to the HisTag on CsgA subunit. (The engineered SpyCatcher was used for future pure hydrogenase binding.) The solution was centrifuged and the sediments contained biofilm beads covered with NR. This sediment was resuspended in PBS and was added to the reaction solution consisting of <em>E. coli</em> with engineered hydrogenase (wet weight 100ug) resuspended in PBS, 150Mm NaCl, 100mM VitaminC, and mediator solution (5mM Paraquat dichloride, for mediating the electrons across the cell membrane). The whole solution including bacteria is adjusted to pH=4 by 100mM Tris-HCl(pH=7.0), given that the pH of 4 was reported to be an optimal environment.<span style=”font-size:12px”> </span> Prior to the assay, the <em>E. coli</em> was induced with IPTG overnight at room temperature. | ||
| − | + | In the activity assay of the hydrogenase in producing hydrogen, the system goes through three periods of “light-on and light-off”. The result (see below) shows the stability of the system and the reversible catalytic activity of the hydrogenase of the reaction, 2H+ + 2e- ⇿ H2 .<p></p> | |
| − | + | <h2> Instrument</h2> | |
<div class="col-lg-6"> | <div class="col-lg-6"> | ||
Revision as of 10:40, 19 October 2016
