Difference between revisions of "Team:CCU Taiwan/Safety"

 
(33 intermediate revisions by 6 users not shown)
Line 6: Line 6:
  
 
<meta name="viewport" content="width=device-width, initial-scale=1">
 
<meta name="viewport" content="width=device-width, initial-scale=1">
−
<script type="text/javascript" src="https://2018.igem.org/Template:CCU_Taiwan/Javascript/ScrollIndicator?
 
−
action=raw&ctype=text/javascript"></script>
 
−
<script type="text/javascript" src="https://2018.igem.org/Template:CCU_Taiwan/Javascript/Scrolling_Parallax?
 
−
action=raw&ctype=text/javascript"></script>
 
−
<link rel="stylesheet"
 
−
  href="https://cdnjs.cloudflare.com/ajax/libs/animate.css/3.5.2/animate.min.css">
 
−
<script type="text/javascript" src="https://2018.igem.org/Template:CCU_Taiwan/Javascript/skrollr?
 
−
action=raw&ctype=text/javascript"></script>
 
 
<script>
 
<script>
 
$(document).ready(function(){
 
$(document).ready(function(){
Line 48: Line 40:
 
     $(this).css("background","black")
 
     $(this).css("background","black")
 
},function(){
 
},function(){
−
     $(this).css("background","#842b00")
+
     $(this).css("background","#31a97e")
 
   });
 
   });
−
 
−
$("#home1").hover(function(){
 
−
$("#home_img").css('display', 'block');
 
−
$("#home_img").animate({ opacity: 0 }, 0);
 
−
$("#home_img").stop(true,false,true).animate({opacity: 1,left:'25vw'},800, 'swing')
 
−
$("#home_word").stop(true,false,true).delay(750).fadeIn(700)
 
−
},function(){
 
−
$("#home_img").stop(true,false,true).css("display","none").css("left","40vw");
 
−
$("#home_word").fadeOut(0)
 
−
});
 
−
 
 
});
 
});
  
−
$(window).scroll(function() {
+
// The function actually applying the offset
−
    var wintop = $(window).scrollTop(), docheight =
+
function offsetAnchor() {
 +
  if (location.hash.length !== 0) {
 +
    window.scrollTo(window.scrollX, window.scrollY - 90);
 +
  }
 +
}
  
−
    $(document).height(), winheight = $(window).height();
+
// Captures click events of all <a> elements with href starting with #
−
    var scrolled = (wintop/(docheight-winheight))*100;
+
$(document).on('click', 'a[href^="#"]', function(event) {
 +
  // Click events are captured before hashchanges. Timeout
 +
  // causes offsetAnchor to be called after the page jump.
 +
  window.setTimeout(function() {
 +
    offsetAnchor();
 +
  }, 0);
 +
});
  
−
    $('.scroll-line').css('height', (scrolled*0.5 + '%'));
+
// Set the offset when entering page with hash present in the url
−
});
+
window.setTimeout(offsetAnchor, 0);
 
 
−
var s = skrollr.init({
+
$(document).scroll(function () { // remove "$"
−
render: function(data) {
+
    var y = $(this).scrollTop();       
−
  //Log the current scroll position.
+
    if (y > 300) {
−
  $('#info').text(data.curTop);
+
        $('.indicator').fadeIn();
−
}
+
    }
−
});;
+
    else {
 +
        $('.indicator').fadeOut();
 +
    }
 +
});
 
 
 
</script>
 
</script>
Line 85: Line 79:
 
<body>
 
<body>
 
   <header>
 
   <header>
−
         <nav class="navbar navbar-default navbar-inverse navbar-fixed-top">
+
         <nav>
 
             <div class="container">
 
             <div class="container">
 
                 <ul class="front">
 
                 <ul class="front">
Line 92: Line 86:
 
'https://2018.igem.org/Team:CCU_Taiwan';"> <img src="https://static.igem.org/mediawiki/2018/0/08/T--CCU_Taiwan--home_button.png"></img></div>
 
'https://2018.igem.org/Team:CCU_Taiwan';"> <img src="https://static.igem.org/mediawiki/2018/0/08/T--CCU_Taiwan--home_button.png"></img></div>
  
−
                     <li class="title" style="cursor:pointer;" id="Home"><img class="img_title" src="https://static.igem.org/mediawiki/2018/2/24/T--CCU_Taiwan--aboutus.png"></img><a>About Us</a><hr>
+
                     <li class="title" style="cursor:pointer;" id="Home"><img class="img_title" src="https://static.igem.org/mediawiki/2018/2/24/T--CCU_Taiwan--aboutus.png"></img><a>About Us</a>
 
                         <ul class="sub" id="sub_home" style="cursor:default;">
 
                         <ul class="sub" id="sub_home" style="cursor:default;">
−
                             <li class="list" id="home1"><a href="https://2018.igem.org/Team:CCU_Taiwan/Team">Team</a></li>
+
                             <a href="https://2018.igem.org/Team:CCU_Taiwan/Team"><li class="list" id="home1">Team</li></a>
−
<li class="list" id="home2"><a href="https://2018.igem.org/Team:CCU_Taiwan/Attributions">Attributions</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Attributions"><li class="list" id="home2">Attributions</li></a>
−
<li class="list" id="home2"><a href="https://2018.igem.org/Team:CCU_Taiwan/Medal">Medal</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Medal"><li class="list" id="home3">Medals</li></a>
−
<li class="list" id="home3"><a href="https://2018.igem.org/Team:CCU_Taiwan/Judge">For Judge</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Judge"><li class="list" id="home4">For Judges</li></a>
−
<hr>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Achievements"><li class="list" id="home5">Achievements</li></a>
−
<div id="home_img">
+
−
<img src="https://static.igem.org/mediawiki/2018/5/57/T--CCU_Taiwan--home.png"></img>
+
−
</div>
+
−
<div id="home_word">
+
−
<a>There's a home for us, always.</a>
+
−
</div>
+
 
                         </ul>
 
                         </ul>
 
                     </li>
 
                     </li>
−
                     <li class="title" style="cursor:pointer;" id="Project"><img class="img_title" src="https://static.igem.org/mediawiki/2018/6/6f/T--CCU_Taiwan--project.png"></img><a>Project</a><hr>
+
                     <li class="title" style="cursor:pointer;" id="Project"><img class="img_title" src="https://static.igem.org/mediawiki/2018/6/6f/T--CCU_Taiwan--project.png"></img><a>Project</a>
 
<ul class="sub" id="sub_project" style="cursor:default;">
 
<ul class="sub" id="sub_project" style="cursor:default;">
−
    <li class="list" id="project1"><a href="https://2018.igem.org/Team:CCU_Taiwan/Description">Description</a></li>
+
    <a href="https://2018.igem.org/Team:CCU_Taiwan/Description"><li class="list" id="project1">Description</li></a>
−
<li class="list" id="project2"><a href="https://2018.igem.org/Team:CCU_Taiwan/Design">Design</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Applied_Design"><li class="list" id="project2">Design</li></a>
−
<li class="list" id="project3"><a href="https://2018.igem.org/Team:CCU_Taiwan/Results">Results</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Results"><li class="list" id="project3">Results</li></a>
−
<li class="list" id="project6"><a href="https://2018.igem.org/Team:CCU_Taiwan/Demonstrate">Demonstration</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Demonstrate"><li class="list" id="project4">Demonstration</li></a>
−
<li class="list" id="project5"><a href="https://2018.igem.org/Team:CCU_Taiwan/InterLab ">InterLab</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/InterLab"><li class="list" id="project5">InterLab</li></a>
−
<hr>
+
 
</ul>
 
</ul>
 
</li>
 
</li>
−
                     <li class="title" style="cursor:pointer;" id="Parts"><img class="img_title" src="https://static.igem.org/mediawiki/2018/1/17/T--CCU_Taiwan--part.png"></img><a>Parts</a><hr>
+
                     <li class="title" style="cursor:pointer;" id="Parts"><img class="img_title" src="https://static.igem.org/mediawiki/2018/1/17/T--CCU_Taiwan--part.png"></img><a>Parts</a>
 
                         <ul class="sub" id="sub_parts" style="cursor:default;">
 
                         <ul class="sub" id="sub_parts" style="cursor:default;">
−
    <li class="list" id="parts1"><a href="https://2018.igem.org/Team:CCU_Taiwan/Parts">Overview</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Parts"><li class="list" id="parts1">Overview</li></a>
−
<li class="list" id="parts2"><a href="https://2018.igem.org/Team:CCU_Taiwan/Basic_Part">Basic Part</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Basic_Part"><li class="list" id="parts1">Basic Part</li></a>
−
<li class="list" id="parts3"><a href="https://2018.igem.org/Team:CCU_Taiwan/Composite_Part">Composite Part</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Composite_Part"><li class="list" id="parts2">Composite Part</li></a>
−
<li class="list" id="parts4"><a href="https://2018.igem.org/Team:CCU_Taiwan/Improve">Improved Part</a></li>
+
−
<hr>
+
 
                         </ul>
 
                         </ul>
 
                     </li>
 
                     </li>
−
                     <li class="title" style="cursor:pointer;" id="Modeling"><img class="img_title" src="https://static.igem.org/mediawiki/2018/0/09/T--CCU_Taiwan--model.png"></img><a>Modeling</a><hr>
+
                     <li class="title" style="cursor:pointer;" id="Modeling"><img class="img_title" src="https://static.igem.org/mediawiki/2018/0/09/T--CCU_Taiwan--model.png"></img><a>Modeling</a>
 
                         <ul class="sub" id="sub_modeling" style="cursor:default;">
 
                         <ul class="sub" id="sub_modeling" style="cursor:default;">
−
                             <li class="list" id="model1"><a href="https://2018.igem.org/Team:CCU_Taiwan/Model">Overview</a></li>
+
                             <a href="https://2018.igem.org/Team:CCU_Taiwan/Model"><li class="list" id="model1">Overview</li></a>
−
                            <li class="list" id="model2"><a href="https://2018.igem.org/Team:CCU_Taiwan/Enzyme">Enzyme Model</a></li>
+
                            <a href="https://2018.igem.org/Team:CCU_Taiwan/Binding"><li class="list" id="model2">Binding Model</li></a>
−
<li class="list" id="model3"><a href="https://2018.igem.org/Team:CCU_Taiwan/Lignin">Lignin Model</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Polymer"><li class="list" id="model3">Polymer Model</li></a>
−
<li class="list" id="model4"><a href="https://2018.igem.org/Team:CCU_Taiwan/Tools">Tools</a></li>
+
−
<hr>
+
 
                         </ul>
 
                         </ul>
 
                     </li>
 
                     </li>
−
                     <li class="title" style="cursor:pointer;" id="Drylab"><img class="img_title" src="https://static.igem.org/mediawiki/2018/f/fc/T--CCU_Taiwan--Dry_lab.png"></img><a>Drylab</a><hr>
+
                     <li class="title" style="cursor:pointer;" id="Drylab"><img class="img_title" src="https://static.igem.org/mediawiki/2018/f/fc/T--CCU_Taiwan--Dry_lab.png"></img><a>Product</a>
 
                         <ul class="sub" id="sub_drylab" style="cursor:default;">
 
                         <ul class="sub" id="sub_drylab" style="cursor:default;">
−
                             <li class="list" id="drylab1"><a href="https://2018.igem.org/Team:CCU_Taiwan/Our_Plan">Overview</a></li>
+
                             <a href="https://2018.igem.org/Team:CCU_Taiwan/Our_Plan"><li class="list" id="drylab1">Analysis</li></a>
−
<li class="list" id="drylab2"><a href="https://2018.igem.org/Team:CCU_Taiwan/Engineering">Engineering</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Engineering"><li class="list" id="drylab2">Production Line</li></a>
−
<li class="list" id="drylab3"><a href="https://2018.igem.org/Team:CCU_Taiwan/Safety">Safety</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Safety"><li class="list" id="drylab3">Safety</li></a>
−
<hr>
+
 
                         </ul>
 
                         </ul>
 
                     </li>
 
                     </li>
−
    <li class="title" style="cursor:pointer;" id="Human_Practice"><img class="img_title" src="https://static.igem.org/mediawiki/2018/9/96/T--CCU_Taiwan--humanpractice.png"></img><a>HP</a><hr>
+
    <li class="title" style="cursor:pointer;" id="Human_Practice"><img class="img_title" src="https://static.igem.org/mediawiki/2018/9/96/T--CCU_Taiwan--humanpractice.png"></img><a>HP</a>
 
                         <ul class="sub" id="sub_human_practice" style="cursor:default;">
 
                         <ul class="sub" id="sub_human_practice" style="cursor:default;">
−
                             <li class="list" id="human_practice1"><a href="https://2018.igem.org/Team:CCU_Taiwan/Human_Practices">Overview</a></li>
+
                             <a href="https://2018.igem.org/Team:CCU_Taiwan/Human_Practices"><li class="list" id="human_practice1">Human Practice</li></a>
−
<li class="list" id="human_practice2"><a href="https://2018.igem.org/Team:CCU_Taiwan/Engagement">Public</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Public_Engagement"><li class="list" id="human_practice2">Public Engagement</li></a>
−
<li class="list" id="human_practice3"><a href="https://2018.igem.org/Team:CCU_Taiwan/Entrepreneurship">Entrepreneurship</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Entrepreneurship"><li class="list" id="human_practice3">Entrepreneurship</li></a>
−
<li class="list" id="human_practice4"><a href="https://2018.igem.org/Team:CCU_Taiwan/Intergrate">Intergrate</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/engaging_experts"><li class="list" id="human_practice4">Engaging Experts</li></a>
−
<hr>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Integrate"><li class="list" id="human_practice5">Integrated HP</li></a>
 
                         </ul>
 
                         </ul>
 
                     </li>
 
                     </li>
 
    <li class="title" style="cursor:pointer;" id="Notebook"><img class="img_title" src="https://static.igem.org/mediawiki/2018/c/c9/T--CCU_Taiwan--notebook.png"></img><a>Notebook</a>
 
    <li class="title" style="cursor:pointer;" id="Notebook"><img class="img_title" src="https://static.igem.org/mediawiki/2018/c/c9/T--CCU_Taiwan--notebook.png"></img><a>Notebook</a>
 
                         <ul class="sub" id="sub_notebook" style="cursor:default;">
 
                         <ul class="sub" id="sub_notebook" style="cursor:default;">
−
                             <li class="list" id="notebook1"><a href="https://2018.igem.org/Team:CCU_Taiwan/Notebook">Overview</a></li>
+
                             <a href="https://2018.igem.org/Team:CCU_Taiwan/Notebook"><li class="list" id="notebook1">Overview</li></a>
−
<li class="list" id="notebook2"><a href="https://2018.igem.org/Team:CCU_Taiwan/Collaborations">Collaborations</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Collaborations"><li class="list" id="notebook2">Collaborations</li></a>
−
<li class="list" id="notebook3"><a href="https://2018.igem.org/Team:CCU_Taiwan/Experiments">Experiments</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Protocols"><li class="list" id="notebook3">Protocols</li></a>
−
<li class="list" id="notebook4"><a href="https://2018.igem.org/Team:CCU_Taiwan/DATABASE">DATABASE</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Experiments"><li class="list" id="notebook4">Experiments</li></a>
−
<hr>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Materials"><li class="list" id="notebook5">Materials</li></a>
 
                         </ul>
 
                         </ul>
 
                     </li>
 
                     </li>
Line 164: Line 146:
 
             </div>
 
             </div>
 
         </nav>
 
         </nav>
−
<!--<div class="scroll-line"></div>
+
 
−
<div class="scroll-line-bg"></div>
+
−
<div class="Quick-navigation">
+
−
  <a href="#A" class="Quick-navigation-item">____Background</a>
+
−
  <a href="#B" class="Quick-navigation-item">____Calibration 1</a>
+
−
  <a href="#C" class="Quick-navigation-item">____Calibration 2</a>
+
−
  <a href="#D" class="Quick-navigation-item">____Calibration 3</a>
+
−
  <a href="#E" class="Quick-navigation-item">____Calibration 4</a>
+
−
  <a href="#F" class="Quick-navigation-item">____Calibration 5</a>
+
−
  <div class="Scroll-progress-indicator"></div>
+
−
</div>-->
+
 
     </header>
 
     </header>
−
<div class="background">
+
<div class="indicator">
−
<div class="bcg"></div>
+
 
−
      <div class="content">
+
<div class="pointerDrylab" id="1"><a href="#ca1">Filtering system</a></div>
−
<p class="first">Safety</p>
+
<div class="pointerDrylab" id="2"><a href="#ca2">Heating system</a></div>
−
<p class="second">All members</p>
+
 
−
<p class="description">All of our team members have basic experimental operation exercises and have a basic understanding of the experiment. In addition, all team members attended experimental safety lectures held by our school, we all have certain concepts for laboratory safety.</p>
+
</div>
−
<p class="second">Wet Lab</p>
+
 
−
<p class="description">In the biological experiment part, the methanolic yeast and Escherichia coli were used, and the experimental contents were all in the first level of Biosafety Level l lab. The operation sites were all in the aseptic operation table, and the personnel were in the experiment. Gloves were worn during the period and equipment were all disinfected with alcohol before and after the experiment.</p>
+
<div class="backgroundDrylab">
−
<p class="second">Dry Lab</p>
+
<div class="photoSafety"><h1 class="bigtitle">SAFETY<h1></div>
−
<p class="description">In the dry experimental group, we conducted experiments that adhered to laboratory safety rules (such as wearing gloves, using volatile drugs in the draft cabinet, etc.) and having relevant professional guidance. Before using the instrument, the yeast will be filtered out in the laboratory to prevent the yeast from leaving the laboratory. All reactions in the design of the future production line will be carry out in the fermentation tank. Keep the reaction environment in a sealed state, then filtering out the polymer. By two protective measures, filter and heat, we can prevent any leakage. For the filter, we will be using a 33kDa filter, the yeast will not pass. As for the heater, all products will be heated 110 ° C for 10 minutes.</p>
+
<div class="content">
−
+
<br><br>
−
      </div>
+
<p class="description">&emsp;&emsp;The escaping of genetically modified organisms from labs is a serious problem since it will bring up unpredictable impacts to our ecosystem. We take biosafety very seriously when we design  every parts of our experiment and the design of our production line.
 +
Our design of our production line insure that there's no genetically modified yeasts (also non-genetically modified yeasts) could survived and pass the whole production line and leak into the environment or residue on our products.
 +
We also dealt cautiously with wastes produced from the experiment and participated in safety training, to minimum the chance of causing biohazard pollution, or simply hurt ourselves.
 +
</p>
 +
<p class="first" id="ca1">Filtering system</p>
 +
<p class="description">&emsp;&emsp;The average size of the <I>P. pastoris</I> is about 4–6 μm (Gmeiner, C. et al. 2015), we use the MCE filter to separate the yeasts from leaking out the fermentation tank.
 +
</p>
 +
<p class="first" id="ca2">Heating system</p>
 +
<p class="description">&emsp;&emsp;According to the literature (Martínez, D. et al. 2015), heating at 70 °C for 1 day or incubate at 30 °C for 48 hr, is sufficient to kill all the yeasts.
 +
The minimum operating temperature of extrusion granulation will be over 110 °C. Therefore, no yeast could survived in the production line.
 +
</p>
 +
<p class="description">Following are our heating tests:</p>
 +
                <div id="Safety1" class="polaroid" style="display:inline-block">
 +
                  <img src="https://static.igem.org/mediawiki/2018/e/e8/T--CCU_Taiwan--CCUsafety.jpg" width="100%">
 +
                  <div class="container">
 +
                    <p>Figure1: <I>P. pastoris</I> yeast heated at 30 °C for 1 day , and incubated at 30 °C for 48 hr.<br>(left: 0.005, right: 0.05)</p>
 +
                  </div>
 +
                </div>
 +
                <div id="Safety2" class="polaroid" style="display:inline-block">
 +
                  <img src="https://static.igem.org/mediawiki/2018/4/4c/T--CCU_Taiwan--safety2.png" width="100%">
 +
                  <div class="container">
 +
                    <p>Figure2: <I>P. pastoris</I> yeast heated at 50 °C for 1 day , and incubated at 30 °C for 48 hr.</p>
 +
                  </div>
 +
                </div>
 +
                <div id="Safety3" class="polaroid" style="display:inline-block">
 +
                  <img src="https://static.igem.org/mediawiki/2018/1/1e/T--CCU_Taiwan--safety3.png" width="100%">
 +
                  <div class="container">
 +
                    <p>Figure3: <I>P. pastoris</I> yeast heated at 70 °C for 1 day , and incubate at 30 °C for 48 hr.</p>
 +
                  </div>
 +
                </div><br><br>
 +
<p class="description">&emsp;&emsp;We found that our <I>P. pastoris</I> diden't survive after were heated at 50 °C for 30 minutes. Which proof that the heat our production line generated could kill <I>P. pastoris</I> which pass the filter.</p>
 +
 +
<br>
 +
<p class="second"><br>Reference</p>
 +
<p class="description">
 +
Gmeiner, C., Saadati, A., Maresch, D., Krasteva, S., Frank, M., Altmann, F., … Spadiut, O. (2015). Development of a fed-batch process for a recombinant <I>Pichia pastoris</I> Δoch1 strain expressing a plant peroxidase. Microbial Cell Factories, 14(1). doi:10.1186/s12934-014-0183-3<br>
 +
<br>Bacher, G., Szymanski, W. W., Kaufman, S. L., Zöllner, P., Blaas, D., & Allmaier, G. (2001). Charge-reduced nano electrospray ionization combined with differential mobility analysis of peptides, proteins, glycoproteins, noncovalent protein complexes and viruses. Journal of Mass Spectrometry, 36(9), 1038–1052.doi:10.1002/jms.208<br>
 +
<br>3. Martínez, D., Menéndez, C., Echemendia, F. M., Hernández, L., Sobrino, A., & Trujillo, L. E. (2015). Kinetics of sucrose hydrolysis by immobilized recombinant <I>Pichia pastoris</I> cells in a batch reactors. J Microb Biochem Technol, 7, 294-6.<br>
 +
 +
</p>
 +
<br><br><br>
 +
</div>
 
</div>
 
</div>
 
</body>
 
</body>
 +
<footer class="footer">
 +
            <a href="http://huitong.com.tw">
 +
              <img  id="BSlogo" src="https://static.igem.org/mediawiki/2018/0/07/T--CCU_Taiwan--CCUP1.png" align="bottom">
 +
            </a>
 +
 +
            <a href="https://www.facebook.com/ccuigemteam/?ref=br_rs">
 +
            <img  id="CCUTEAMlogo" src="https://static.igem.org/mediawiki/2018/e/e3/T--CCU_Taiwan--CCUP2.png" align="bottom">
 +
            </a>
  
−
</html>
+
            <a href="http://ewww.ccu.edu.tw/">
 +
            <img  id="CCUlogo" src="https://static.igem.org/mediawiki/2018/8/89/T--CCU_Taiwan--CCUP3.png" align="bottom">
 +
            </a>
 +
</footer>

Latest revision as of 08:50, 1 December 2018

SAFETY



  The escaping of genetically modified organisms from labs is a serious problem since it will bring up unpredictable impacts to our ecosystem. We take biosafety very seriously when we design every parts of our experiment and the design of our production line. Our design of our production line insure that there's no genetically modified yeasts (also non-genetically modified yeasts) could survived and pass the whole production line and leak into the environment or residue on our products. We also dealt cautiously with wastes produced from the experiment and participated in safety training, to minimum the chance of causing biohazard pollution, or simply hurt ourselves.

Filtering system

  The average size of the P. pastoris is about 4–6 μm (Gmeiner, C. et al. 2015), we use the MCE filter to separate the yeasts from leaking out the fermentation tank.

Heating system

  According to the literature (Martínez, D. et al. 2015), heating at 70 °C for 1 day or incubate at 30 °C for 48 hr, is sufficient to kill all the yeasts. The minimum operating temperature of extrusion granulation will be over 110 °C. Therefore, no yeast could survived in the production line.

Following are our heating tests:

Figure1: P. pastoris yeast heated at 30 °C for 1 day , and incubated at 30 °C for 48 hr.
(left: 0.005, right: 0.05)

Figure2: P. pastoris yeast heated at 50 °C for 1 day , and incubated at 30 °C for 48 hr.

Figure3: P. pastoris yeast heated at 70 °C for 1 day , and incubate at 30 °C for 48 hr.



  We found that our P. pastoris diden't survive after were heated at 50 °C for 30 minutes. Which proof that the heat our production line generated could kill P. pastoris which pass the filter.



Reference

Gmeiner, C., Saadati, A., Maresch, D., Krasteva, S., Frank, M., Altmann, F., … Spadiut, O. (2015). Development of a fed-batch process for a recombinant Pichia pastoris Δoch1 strain expressing a plant peroxidase. Microbial Cell Factories, 14(1). doi:10.1186/s12934-014-0183-3

Bacher, G., Szymanski, W. W., Kaufman, S. L., Zöllner, P., Blaas, D., & Allmaier, G. (2001). Charge-reduced nano electrospray ionization combined with differential mobility analysis of peptides, proteins, glycoproteins, noncovalent protein complexes and viruses. Journal of Mass Spectrometry, 36(9), 1038–1052.doi:10.1002/jms.208

3. Martínez, D., Menéndez, C., Echemendia, F. M., Hernández, L., Sobrino, A., & Trujillo, L. E. (2015). Kinetics of sucrose hydrolysis by immobilized recombinant Pichia pastoris cells in a batch reactors. J Microb Biochem Technol, 7, 294-6.