Difference between revisions of "Team:BJRS China/Design"

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<title>BJRS</title>
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<h1>Design</h1>
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Design is the first step in the design-build-test cycle in engineering and synthetic biology. Use this page to describe the process that you used in the design of your parts. You should clearly explain the engineering principles used to design your project.
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  <div class="top container-fluid" style="position:fixed; z-index:999; top:0;">
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<div class="navbar-header"><a href="https://2018.igem.org/Team:BJRS_China" target="_top" class="navbar-brand"><img src="https://static.igem.org/mediawiki/2018/7/73/T--BJRS_China--logo.png" width="120px" height="120px" style="position:relative;top:0px;left:90%;" class="img-fluid rounded"><img src="https://static.igem.org/mediawiki/2018/b/b3/T--BJRS_China--logo2.jpeg"width="1200px" height="50px"style="position:relative;top:-114px;left:10px;"></a></div>
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    <div>
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      <ul class="nav navbar-nav">
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        <li class="dropdown"> <a href="#" class="dropdown-toggle" data-toggle="dropdown"> PROJECT <b class="caret"></b> </a>
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          <ul class="dropdown-menu">
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            <li><a href="https://2018.igem.org/Team:BJRS_China/Background">BACKGROUND</a></li>
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<li><a href="https://2018.igem.org/Team:BJRS_China/Design">DESIGN</a></li> 
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            <li><a href="https://2018.igem.org/Team:BJRS_China/Results">RESULTS</a></li>
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            <li><a href="https://2018.igem.org/Team:BJRS_China/Improve">IMPROVE</a></li>
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<li><a href="https://2018.igem.org/Team:BJRS_China/InterLab">INTERLAB</a></li>
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<li><a href="https://2018.igem.org/Team:BJRS_China/Demonstrate">DEMONSTRATE</a></li>
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          </ul>
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        </li>
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        <li><a href="https://2018.igem.org/Team:BJRS_China/Model">MODEL</a></li>
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        <li class="dropdown"> <a href="#" class="dropdown-toggle" data-toggle="dropdown"> HUMAN PRACTICE <b class="caret"></b> </a>
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          <ul class="dropdown-menu">
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            <li><a href="https://2018.igem.org/Team:BJRS_China/Human_Practices">Human Practices</a></li>
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            <li><a href="https://2018.igem.org/Team:BJRS_China/Public_Engagement">Public Engagement</a></li>
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          </ul>
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        </li>
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        <li class="dropdown"> <a href="#" class="dropdown-toggle" data-toggle="dropdown"> ACHIEVEMENTS <b class="caret"></b> </a>
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          <ul class="dropdown-menu">
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            <li><a href="https://2018.igem.org/Team:BJRS_China/Parts">PARTS</a></li>
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            <li><a href="https://2018.igem.org/Team:BJRS_China/Judging_Criteria">JUDGING CRITERIA</a></li>
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          </ul>
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        </li>
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        <li><a href="https://2018.igem.org/Team:BJRS_China/Safety">SAFETY</a></li>
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        <li class="dropdown"> <a href="#" class="dropdown-toggle" data-toggle="dropdown"> ATTRIBUTION <b class="caret"></b> </a>
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          <ul class="dropdown-menu">
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            <li><a href="https://2018.igem.org/Team:BJRS_China/Attributions">ATTRIBUTION</a></li>
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            <li><a href="https://2018.igem.org/Team:BJRS_China/Collaborations">COLLABORATION</a></li>
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          </ul>
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        </li>
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        <li class="dropdown"> <a href="#" class="dropdown-toggle" data-toggle="dropdown"> TEAM <b class="caret"></b> </a>
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          <ul class="dropdown-menu">
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            <li><a href="https://2018.igem.org/Team:BJRS_China/Members">MEMBERS</a></li>
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            <li><a href="https://2018.igem.org/Team:BJRS_China/Notebooks">NOTEBOOKS</a></li>
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<li><a href="https://2018.igem.org/Team:BJRS_China/Protocol">PROTOCOL</a></li>
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<li><a href="https://2018.igem.org/Team:BJRS_China/Acknowledgement">ACKNOWLEDGEMENT</a></li>
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          </ul>
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        </li>
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      </ul>
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    </div>
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</nav>
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This page is different to the "Applied Design Award" page. Please see the <a href="https://2018.igem.org/Team:BJRS_China/Applied_Design">Applied Design</a> page for more information on how to compete for that award.
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          <h1 style = "text-align: center">Design</h1>
  
 
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<div style="padding-left:180px;padding-right:180px;color:black;">
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            <h2>Overview</h2>
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<p style="line-height:30px"><font size="4">&emsp;&emsp;This year our project focused on enhancing the growth and production efficiency of the engineered bacteria during high-density fermentation. Previous research has found that expressing <i>Vitreoscilla</i> hemoglobin (VHb) inside the bacteria could leverage its property of combining and releasing oxygen to improve the efficiency of oxygen utilization and therefore enhance the efficiency of bacteria growth and production.<sup>[1]</sup> Follow-up studies have shown that expressing VHb in the periplasmic space could further improve that efficiency compared with intracellular expression.<sup>[2]</sup> Based on these, we proposed that expressing VHb outside the outer membrane using surface display system could further enhance the oxygen density surrounding the bacteria and improve oxygen utility in the system for further enhancement of the fermentation efficiency.</font></p>
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<figure class="text-center">
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<img src="https://static.igem.org/mediawiki/2018/8/85/T--BJRS_China--design2.jpg" width="60%" >
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            </figure>
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<h2>Surface display system</h2>
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<p style="line-height:30px"><font size="4">&emsp;&emsp;Our project used three different surface display system including INP (<a href='http://parts.igem.org/Part:BBa_K523013'>BBa_K523013</a>), intimin and autotransporter. We fused these surface transporter proteins with fluorescent protein meos and sfGFP respectively and observed the bacteria carrying these plasmids under confocal microscope to confirm that these systems could work effectively.</font></p>
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<figure class="text-center">
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<img src="https://static.igem.org/mediawiki/2018/a/a8/T--BJRS_China--design3.jpg" width="50%" >
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                <figcaption><div style="padding-left:300px;padding-right:300px"><b>Figure 1</b>  The framework of the three kinds of surface displaying fluorescent protein parts.
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              </div> </figcaption>
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            </figure>
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<h2>Growth efficiency</h2>
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<p style="line-height:30px"><font size="4">&emsp;&emsp;We compared the growth efficiency of <i>E.coli</i> BL21 expressing VHb in the cell and on cell surface respectively mainly using the following parts.</font></p>
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<figure class="text-center">
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<img src="https://static.igem.org/mediawiki/parts/d/d1/T--BJRS_China--result6.jpg" width="65%" >
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                <figcaption><div style="padding-left:18%;padding-right:15%"><b>Figure 2</b> Construction of INP-VHb and intracellular expressing VHb vectors.
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                </div></figcaption>
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            </figure>
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<h3>References</h3>
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<p style="line-height:30px"><font size="3">1.Lei Zhang <i>et al.</i>. Recent developments and future prospects of <i>Vitreoscilla</i> hemoglobin application in metabolic engineering. <i>Biotechnology Advances</i> <b>25</b>(2007).</font></p>
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<p style="line-height:30px"><font size="3">2.Pengfei Ouyang <i>et al.</i>. Increasing oxygen availability for improving poly(3-hydroxybutyrate) production by <i>Halomonas</i>. <i>Metabolic Engineering</i> <b>45</b>(2018).</font></p>
  
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<h3>What should this page contain?</h3>
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<li>Explanation of the engineering principles your team used in your design</li>
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<li>Discussion of the design iterations your team went through</li>
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<li>Experimental plan to test your designs</li>
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<h3>Inspiration</h3>
 
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<li><a href="https://2016.igem.org/Team:MIT/Experiments/Promoters">2016 MIT</a></li>
 
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<li><a href="https://2016.igem.org/Team:BostonU/Proof">2016 BostonU</a></li>
 
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<li><a href="https://2016.igem.org/Team:NCTU_Formosa/Design">2016 NCTU Formosa</a></li>
 
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Latest revision as of 23:35, 17 October 2018

BJRS


Design

Overview

  This year our project focused on enhancing the growth and production efficiency of the engineered bacteria during high-density fermentation. Previous research has found that expressing Vitreoscilla hemoglobin (VHb) inside the bacteria could leverage its property of combining and releasing oxygen to improve the efficiency of oxygen utilization and therefore enhance the efficiency of bacteria growth and production.[1] Follow-up studies have shown that expressing VHb in the periplasmic space could further improve that efficiency compared with intracellular expression.[2] Based on these, we proposed that expressing VHb outside the outer membrane using surface display system could further enhance the oxygen density surrounding the bacteria and improve oxygen utility in the system for further enhancement of the fermentation efficiency.


Surface display system

  Our project used three different surface display system including INP (BBa_K523013), intimin and autotransporter. We fused these surface transporter proteins with fluorescent protein meos and sfGFP respectively and observed the bacteria carrying these plasmids under confocal microscope to confirm that these systems could work effectively.

Figure 1 The framework of the three kinds of surface displaying fluorescent protein parts.

Growth efficiency

  We compared the growth efficiency of E.coli BL21 expressing VHb in the cell and on cell surface respectively mainly using the following parts.

Figure 2 Construction of INP-VHb and intracellular expressing VHb vectors.

References

1.Lei Zhang et al.. Recent developments and future prospects of Vitreoscilla hemoglobin application in metabolic engineering. Biotechnology Advances 25(2007).

2.Pengfei Ouyang et al.. Increasing oxygen availability for improving poly(3-hydroxybutyrate) production by Halomonas. Metabolic Engineering 45(2018).