LechenQian (Talk | contribs) |
|||
(4 intermediate revisions by 2 users not shown) | |||
Line 13: | Line 13: | ||
<ul id="sidebar" class="nav bs-docs-sidenav "> | <ul id="sidebar" class="nav bs-docs-sidenav "> | ||
<li > | <li > | ||
− | <a href="#p10"> | + | <a href="#p10">Overview</a> |
</li> | </li> | ||
<li> | <li> | ||
− | <a href="#Introduction to Solar Hunter | + | <a href="#Introduction to Solar Hunter">Introduction to Solar Hunter</a> |
</li> | </li> | ||
<li> | <li> | ||
− | <a href="#Demonstrated Functionality" style="font-size:14px | + | <a href="#Demonstrated Functionality" >Demonstrated Functionality</a> |
+ | <ul> | ||
+ | <li> | ||
+ | <a href="#Methods"style="font-size:14px" >Methods</a> | ||
+ | </li> | ||
+ | <li> | ||
+ | <a href="#Instrument" style="font-size:14px">Instrument</a> | ||
+ | </li> | ||
+ | <li> | ||
+ | <a href="#Results"style="font-size:14px" >Results</a> | ||
+ | </li> | ||
+ | </ul> | ||
</li> | </li> | ||
<li> | <li> | ||
− | <a href="#Efficiency | + | <a href="#Efficiency">Efficiency</a> |
</li> | </li> | ||
<li> | <li> | ||
− | <a href="#Potential use for wider application | + | <a href="#Potential use for wider application" >Potential use for wider application</a> |
</li> | </li> | ||
<li> | <li> | ||
− | <a href="#Conclusion | + | <a href="#Conclusion" >Conclusion</a> |
</li> | </li> | ||
</ul> | </ul> | ||
Line 40: | Line 51: | ||
<div class="row"> | <div class="row"> | ||
<div class="col-lg-12"> | <div class="col-lg-12"> | ||
− | <h1 align="center"> | + | <h1 align="center">Overview</h1> |
<p> Given that our integrative system has been proved to produce hydrogen with great stability and repeatability and its intrinsic characteristics for scalable production, we think our project is a successful applied design. Here, we summarize the qualification and merits of our design in the following figure. In addition, biofilm-interfaced Nanorods has the potential for easy recycling and is cheap for production, thus presenting an innovative and interesting example towards industrial-oriented artificial photosynthesis system. </p> | <p> Given that our integrative system has been proved to produce hydrogen with great stability and repeatability and its intrinsic characteristics for scalable production, we think our project is a successful applied design. Here, we summarize the qualification and merits of our design in the following figure. In addition, biofilm-interfaced Nanorods has the potential for easy recycling and is cheap for production, thus presenting an innovative and interesting example towards industrial-oriented artificial photosynthesis system. </p> | ||
<center> | <center> | ||
− | <img src="https://static.igem.org/mediawiki/ | + | <img src="https://static.igem.org/mediawiki/2016/0/09/T--ShanghaitechChina--hahahaNEW.jpg" width="80%"> |
</center> | </center> | ||
<p></p><p></p> | <p></p><p></p> | ||
Line 52: | Line 63: | ||
− | <p id=" | + | <p id="Introduction to Solar Hunter"></p> |
<div class="content" > | <div class="content" > | ||
<div class="row"> | <div class="row"> | ||
Line 70: | Line 81: | ||
− | <p id=" | + | <p id="Demonstrated Functionality"></p> |
<div class="content"> | <div class="content"> | ||
<div class="row"> | <div class="row"> | ||
− | <p id=" | + | <p id="Methods"></p> |
<div class="col-lg-12"> <center> <h1 >Demonstrated Functionality</h1></center> | <div class="col-lg-12"> <center> <h1 >Demonstrated Functionality</h1></center> | ||
<h2> Methods </h2> | <h2> Methods </h2> | ||
Line 79: | Line 90: | ||
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> | 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> | ||
− | + | <p id="Instrument" style="margin-bottom:80px"></p> | |
<h2> Instrument</h2> | <h2> Instrument</h2> | ||
Line 91: | Line 102: | ||
<center><p style="text-align:center"><b>Figure 1</b> Apparatus of the hydrogen production assay.</p></center> | <center><p style="text-align:center"><b>Figure 1</b> Apparatus of the hydrogen production assay.</p></center> | ||
It contains (1) an anaerobic reaction container which is a transparent circular cuvette that allows light to go through; (2) a light source in our hydrogen production assay acting as a substitute for the real sun. (We chose a high-power white LED light, set 28cm away from the reaction container for a even distribution of photons); (3) a hydrogen electrode linked to its inner sensor inserted into the reaction container to measure the realtime concentration of hydrogen; (4) a data hub; (5) a computer connected to the hub to record the data and generate the curve of concentration variation within a period of time. <p></p> | It contains (1) an anaerobic reaction container which is a transparent circular cuvette that allows light to go through; (2) a light source in our hydrogen production assay acting as a substitute for the real sun. (We chose a high-power white LED light, set 28cm away from the reaction container for a even distribution of photons); (3) a hydrogen electrode linked to its inner sensor inserted into the reaction container to measure the realtime concentration of hydrogen; (4) a data hub; (5) a computer connected to the hub to record the data and generate the curve of concentration variation within a period of time. <p></p> | ||
− | + | <p id="Results" style="margin-bottom:80px"></p> | |
<h2> Results </h2> | <h2> Results </h2> | ||
Line 122: | Line 133: | ||
<p style="text-align:center"><b>Figure Standard</b> Relationship between voltage data and concentration.</p> | <p style="text-align:center"><b>Figure Standard</b> Relationship between voltage data and concentration.</p> | ||
− | Following the method above , we obtain the rate of hydrogen evolution: the tip of the first period is 7.061 mV at 500s. This corresponds to 2.179 (0.3086*7.061) umol/L at 500s. Thus the rate is 0.0126 (2.179/500*3mL*1000) umol/s, for 0.1g E. | + | Following the method above , we obtain the rate of hydrogen evolution: the tip of the first period is 7.061 mV at 500s. This corresponds to 2.179 (0.3086*7.061) umol/L at 500s. Thus the rate is 0.0126 (2.179/500*3mL*1000) umol/s, for 0.1g <i>E. coli</i>. In comparison with the rate from reference 1, 0.0086mol umol/s. This 46% increase in the efficiency shows that our system not only functions, but is also a big improvement compared with a artificial hydrogen production system reported before .<p></p> |
</div></div></div> | </div></div></div> | ||
Line 148: | Line 159: | ||
<h4><b>Characterization</b></h4> | <h4><b>Characterization</b></h4> | ||
− | As Figure3 illustrated, His-CsgA-SpyCatcher-Histag mutant incubated with mCherry-SpyTag show a clear biofilm-associated mcherry fluorescence signal, which indicating the accurate conformation and function of the SpyTag and SpyCatcher linkage system. The third figure is merged by the first and second figures of each sample are snapped respectively under green laser field with 558 nm wavelength and bright field of fluorescence microscopy, Zeiss Axio Imager Z2. As for controls, strains secreted CsgA–Histag and ΔCsgA both are unable to specifically attach to SpyTag thus no distinct localization highlight of red fluorescence on E.coli. That to a large extent prove the specificity of our desired linkage between SpyTag and SpyCatcher system. <p></p> | + | As Figure3 illustrated, His-CsgA-SpyCatcher-Histag mutant incubated with mCherry-SpyTag show a clear biofilm-associated mcherry fluorescence signal, which indicating the accurate conformation and function of the SpyTag and SpyCatcher linkage system. The third figure is merged by the first and second figures of each sample are snapped respectively under green laser field with 558 nm wavelength and bright field of fluorescence microscopy, Zeiss Axio Imager Z2. As for controls, strains secreted CsgA–Histag and ΔCsgA both are unable to specifically attach to SpyTag thus no distinct localization highlight of red fluorescence on <i>E. coli</i>. That to a large extent prove the specificity of our desired linkage between SpyTag and SpyCatcher system. <p></p> |
<center> | <center> | ||
<img src="https://static.igem.org/mediawiki/parts/5/5c/Shanghaitechchina_mcherry-SpyTag%2BCsgA-SpyCatcher.png" style="width:100%;"> | <img src="https://static.igem.org/mediawiki/parts/5/5c/Shanghaitechchina_mcherry-SpyTag%2BCsgA-SpyCatcher.png" style="width:100%;"> | ||
Line 162: | Line 173: | ||
<div class="col-lg-12" > | <div class="col-lg-12" > | ||
<h1 align="center" >Conclusion</h1> | <h1 align="center" >Conclusion</h1> | ||
− | In conclusion, E. | + | In conclusion, <i>E. coli</i> strains expressing biofilm on microspheres to anchor nanorods and strains expressing hydrogenase work great together for producing hydrogen. The system achieved a fairly good hydrogen production rate compared to a work reported this year, a nearly 50% increase. 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 previous reports using hydrogenase. Practically speaking, the system comprising <i>E. coli</i> and biofilms are both amenable for scalable operation, rendering itself a great potential for large-scale industrial applications. 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 addition, our SpyCatcher on the CsgA allows the binding of other proteins that may significantly improve our system. The demonstrated efficiency and stability, along with great potential in scalability, recyclability and versatility makes our system an innovative engineering design with potential for industrial application. |
Line 177: | Line 188: | ||
<p>[*] Cao Y, Bai X F. Progress in Research of Preparation of Loaded Nano-CdS and H_2 Production by Photocatalytic Decomposition of Water[J]. Imaging Science & Photochemistry, 2009, 27(3):225-232.</p> | <p>[*] Cao Y, Bai X F. Progress in Research of Preparation of Loaded Nano-CdS and H_2 Production by Photocatalytic Decomposition of Water[J]. Imaging Science & Photochemistry, 2009, 27(3):225-232.</p> | ||
[1] Honda Y, Hagiwara H, Ida S, et al. Application to Photocatalytic H2, Production of a Whole-Cell Reaction by Recombinant Escherichia coli, Cells Expressing [FeFe]-Hydrogenase and Maturases Genes[J]. Angewandte Chemie, 2016<p></p> | [1] Honda Y, Hagiwara H, Ida S, et al. Application to Photocatalytic H2, Production of a Whole-Cell Reaction by Recombinant Escherichia coli, Cells Expressing [FeFe]-Hydrogenase and Maturases Genes[J]. Angewandte Chemie, 2016<p></p> | ||
− | [2] Z. Botyanszki, P. K. R. Tay, P. Q. Nguyen, M. G. Nussbaumer, N. S. Joshi, Engineered catalytic biofilms: Site‐specific enzyme immobilization onto E. coli curli nanofibers. Biotechnology and bioengineering 112, 2016-2024 (2015).<p></p> | + | [2] Z. Botyanszki, P. K. R. Tay, P. Q. Nguyen, M. G. Nussbaumer, N. S. Joshi, Engineered catalytic biofilms: Site‐specific enzyme immobilization onto <i>E. coli</i> curli nanofibers. Biotechnology and bioengineering 112, 2016-2024 (2015).<p></p> |
</div> | </div> | ||
</div> | </div> |
Latest revision as of 23:09, 19 October 2016