Difference between revisions of "Team:TJUSLS China/Demonstrate"

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<html>
 
<html>
  
 
<head>
 
<head>
−
     <!-- head -->
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     <meta charset="UTF-8">
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    <link href="https://2016.igem.org/Team:TJUSLS_China/home/home_style?action=raw&amp;ctype=text/css" rel="stylesheet" type="text/css" media="all" />
+
     <title></title>
−
     <!--start slider -->
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     <link rel="stylesheet" href="https://2016.igem.org/Team:TJUSLS_China/home/index_style?action=raw&amp;ctype=text/css">
−
    <link href="https://2016.igem.org/Team:TJUSLS_China/home/home_lrtk?action=raw&amp;ctype=text/css" rel="stylesheet" type="text/css" media="all" />
+
     <script type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/home/index_js_two?action=raw&ctype=text/javascript"></script>
−
     <link rel="stylesheet" href="https://2016.igem.org/Team:TJUSLS_China/home/home_fwslider?action=raw&amp;ctype=text/css" media="all">
+
     <script type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/home/index_js_one?action=raw&ctype=text/javascript"></script>
−
     <script language="javascript" type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/home/home_lrtk_js?action=raw&ctype=text/javascript"></script>
+
</head>
−
     <script language="javascript" type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/home/home_jquery_min?action=raw&ctype=text/javascript"></script>
+
−
    <script language="javascript" type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/home/home_jquery_min?action=raw&ctype=text/javascript"></script>
+
−
    <script language="javascript" type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/home/home_jquery-ui_min?action=raw&ctype=text/javascript"></script>
+
−
    <script language="javascript" type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/home/home_fwslider1?action=raw&ctype=text/javascript"></script>
+
−
    <!--end slider -->
+
−
    <script language="javascript" type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/home/home_jquery-hover-effect?action=raw&ctype=text/javascript"></script>
+
−
    <script type="text/javascript">
+
−
        //Image Hover
+
−
        jQuery(document).ready(function(){
+
−
            jQuery(function() {
+
−
                jQuery('ul.da-thumbs > li').hoverdir();
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−
            });
+
−
        });
+
−
    </script>
+
−
    <!-- Add fancyBox main JS and CSS files -->
+
−
    <script src="https://2016.igem.org/Team:TJUSLS_China/home/home_jquery.magnific-popup?action=raw&ctype=text/javascript" type="text/javascript"></script>
+
−
    <link href="https://2016.igem.org/Team:TJUSLS_China/home/home_magnific-popup?action=raw&amp;ctype=text/css" rel="stylesheet" type="text/css">
+
−
    <script>
+
−
        $(document).ready(function() {
+
−
            $('.popup-with-zoom-anim').magnificPopup({
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−
                type: 'inline',
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−
                fixedContentPos: false,
+
−
                fixedBgPos: true,
+
−
                overflowY: 'auto',
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−
                closeBtnInside: true,
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−
                preloader: false,
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−
                midClick: true,
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−
                removalDelay: 300,
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                mainClass: 'my-mfp-zoom-in'
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            });
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−
        });
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    </script>
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−
    <!--nav-->
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    <script>
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−
        $(function() {
+
−
            var pull = $('#pull');
+
−
            menu = $('nav ul');
+
−
            menuHeight = menu.height();
+
  
−
            $(pull).on('click', function(e) {
 
−
                e.preventDefault();
 
−
                menu.slideToggle();
 
−
            });
 
−
 
−
            $(window).resize(function(){
 
−
                var w = $(window).width();
 
−
                if(w > 320 && menu.is(':hidden')) {
 
−
                    menu.removeAttr('style');
 
−
                }
 
−
            });
 
−
        });
 
−
    </script>
 
−
 
−
    <!-- origin -->
 
−
    <script>
 
−
        buttonClick = {};
 
−
    </script>
 
−
    <title>Home</title>
 
−
    <!--<script src="js/jquery.min.js"></script>-->
 
−
    <link href="https://2016.igem.org/Team:TJUSLS_China/CSS_Home_bootstr?action=raw&amp;ctype=text/css" rel="stylesheet" type="text/css" media="all">
 
−
    <link href="https://2016.igem.org/Team:TJUSLS_China/CSS_Home_style?action=raw&amp;ctype=text/css" rel="stylesheet" type="text/css" media="all" />
 
−
    <meta name="viewport" content="width=device-width, initial-scale=1">
 
−
    <meta http-equiv="Content-Type" content="text/html; charset=utf-8" />
 
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    <meta name="keywords" content="Solutem  Responsive web template, Bootstrap Web Templates, Flat Web Templates, Andriod Compatible web template,
 
−
Smartphone Compatible web template, free webdesigns for Nokia, Samsung, LG, SonyErricsson, Motorola web design" />
 
−
    <script type="application/x-javascript"> addEventListener("load", function() { setTimeout(hideURLbar, 0); }, false); function hideURLbar(){ window.scrollTo(0,1); } </script>
 
−
    <script language="javascript" type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/js/jquery-min?
 
−
action=raw&ctype=text/javascript"></script>
 
−
    <!---- start-smoth-scrolling---->
 
−
    <script language="javascript" type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/js/move-top?
 
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action=raw&ctype=text/javascript"></script>
 
−
    <script language="javascript" type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/js/easing?
 
−
action=raw&ctype=text/javascript"></script>
 
−
    <script type="text/javascript">
 
−
        jQuery(document).ready(function($) {
 
−
            $(".scroll").click(function(event){
 
−
                event.preventDefault();
 
−
                $('html,body').animate({scrollTop:$(this.hash).offset().top},1200);
 
−
            });
 
−
        });
 
−
    </script>
 
−
    <!---End-smoth-scrolling---->
 
−
    <link href="https://2016.igem.org/Team:TJUSLS_China/CSS_Home_component?action=raw&amp;ctype=text/css" rel="stylesheet" type="text/css"  />
 
−
    <script language="javascript" type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/js/modernizr.custom?
 
−
action=raw&ctype=text/javascript"></script>
 
−
</head>
 
 
<style>
 
<style>
−
    #bodyContent h2 {
 
−
        font-size: 2em;
 
−
    }
 
  
−
     .cbp-spmenu-push-toleft {
+
     img{
−
         left: -115px;
+
         margin-left: 25%;
 
     }
 
     }
  
−
     #content {
+
     #content{width:100%;padding:0px;margin:0px;background-color: #ededed;}
−
        width: 100%;
+
−
        padding: 15px;
+
−
        border: none;
+
−
        color: black;
+
−
        margin-left: 20px;
+
−
        margin-right: auto;
+
−
        background-color: #ffffff;
+
−
        position: relative;
+
−
    }
+
 
     #sideMenu, #top_title
 
     #sideMenu, #top_title
 
     {
 
     {
 
         display: none;
 
         display: none;
 
     }
 
     }
−
 
+
    body{font: 16px sans-serif;}
−
     .right10{
+
     .footer_bg .footer {
−
         right: 10px;
+
         padding: 0;
 +
        border-top: 6px solid #222222;
 
     }
 
     }
−
     .right190{
+
     .footer_bg .footer_btm {
−
         right: 190px;
+
         margin-top: 4%;
 +
        border-top: none;
 +
        padding: 2% 4%;
 +
        padding-top: 2%;
 +
        max-width: 1458px;
 +
        margin: 0 auto;
 
     }
 
     }
−
 
 
</style>
 
</style>
 
<body>
 
<body>
−
<div class="cbp-spmenu-push" id="body">
+
<!--menu-->
−
    <div class="header_bg">
+
<div class="header-box">
−
        <div class="wrap">
+
    <div class="header">
−
            <div class="header">
+
        <!-- start h_menu4 -->
−
                <div class="logo">
+
        <div class="h_menu4">
−
                    <a href="index.html"><img src="images/logo.png" alt="" /></a>
+
            <a class="toggleMenu" href="">Menu</a>
−
                </div>
+
            <ul id="top_ul_my" class="nav">
−
                <!-- start h_menu4 -->
+
                <script>
−
                <div class="h_menu4">
+
                    function getHover(number){
−
                    <a class="toggleMenu" href="">Menu</a>
+
                        var top_ul_my_lis = $("#top_ul_my").find(">li");
−
                    <ul id="top_ul_my" class="nav">
+
                        console.log(top_ul_my_lis);
−
                        <script>
+
                        if(number == 0) top_ul_my_lis[0].setAttribute("class", "active hover");
−
                            function getHover(number){
+
                        else top_ul_my_lis[0].setAttribute("class", "active");
−
                                var top_ul_my_lis = $("#top_ul_my").find(">li");
+
−
                                if(number == 0) top_ul_my_lis[0].setAttribute("class", "active hover");
+
−
                                else top_ul_my_lis[0].setAttribute("class", "active");
+
  
−
                                for(var i = 1; i < top_ul_my_lis.length; i++){
+
                        for(var i = 1; i < top_ul_my_lis.length; i++){
−
                                    if(i == number) {
+
                            if(i == number) {
−
                                        top_ul_my_lis[i].setAttribute("class", "hover");
+
                                top_ul_my_lis[i].setAttribute("class", "hover");
−
                                    }else{
+
                            }else{
−
                                        top_ul_my_lis[i].setAttribute("class", "");
+
                                top_ul_my_lis[i].setAttribute("class", "");
−
                                    }
+
−
                                }
+
 
                             }
 
                             }
 +
                        }
 +
                    }
  
−
                            $(document).ready(function() {
+
                    $(".header").hover(function(event){
−
                                //console.log("ready");
+
                        event.stopPropagation();
−
                                var top_ul_my_lis = $("#top_ul_my").find(">li");
+
                        $('#top_ul_my li').attr("class","");
−
                                for(var i = 1; i < top_ul_my_lis.length; i++) {
+
                    });
−
                                    $(top_ul_my_lis[i]).mouseover(getHover(i));
+
−
                                }
+
−
                            });
+
  
−
                        </script>
+
                </script>
−
                        <li class="active"><a class="active" href="https://2016.igem.org/Team:TJUSLS_China/home">TJUSLS</a></li>
+
                <li onmouseover="getHover(0)"><a href="https://2016.igem.org/Team:TJUSLS_China/home">TJUSLS</a></li>
−
                        <li ><a href="https://2016.igem.org/Team:TJUSLS_China/Project" class="root">project</a>
+
                <li onmouseover="getHover(1)" class=""><a href="" class="root">project</a>
−
                            <ul>
+
                    <ul>
−
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Description">Description</a></li>
+
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Description">Description</a></li>
−
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Design">Design</a></li>
+
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Design">Design</a></li>
−
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Experiments">Experiments</a></li>
+
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Experiments">Experiments</a></li>
−
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Proof">Proof of Concept</a></li>
+
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Proof">Proof of Concept</a></li>
−
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Demonstrate">Demonstrate</a></li>
+
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Demonstrate">Demonstrate</a></li>
−
                               
+
−
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/PROJECT/PROTOCOLS">protocols</a></li>
+
−
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Notebook">Notebook</a></li>
+
−
                            </ul>
+
−
                        </li>
+
−
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Parts">PARTS</a>
+
−
                            <ul>
+
−
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Basic_Part">Basic Parts</a></li>
+
−
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Composite_Part">Composite Parts</a></li>
+
−
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Part_Collection">Part Collection</a></li>
+
  
−
                            </ul>
+
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Notebook">Notebook</a></li>
−
                         </li>
+
                         <li><a href="https://2016.igem.org/Team:TJUSLS_China/Results">Results</a></li>
−
<li><a href="https://2016.igem.org/Team:TJUSLS_China/Modeling">MODELING</a>
+
                    </ul>
−
                            <ul>
+
                </li>
−
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Modeling">Modeling</a></li>
+
                <li onmouseover="getHover(2)" class=""><a href="https://2016.igem.org/Team:TJUSLS_China/Parts">PARTS</a>
−
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Software">Software</a></li>
+
                    <ul>
−
                             
+
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Basic_Part">Basic Parts</a></li>
 +
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Composite_Part">Composite Parts</a></li>
 +
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Part_Collection">Part Collection</a></li>
  
−
                            </ul>
+
                    </ul>
−
                        </li>
+
                </li>
−
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Safety">SAFETY</a></li>
+
                <li class="active" onmouseover="getHover(3)"><a class="active" href="https://2016.igem.org/Team:TJUSLS_China/Safety">SAFETY</a></li>
−
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Attributions">ATTRIBUTIONS</a>
+
                <li onmouseover="getHover(4)" class=""><a href="https://2016.igem.org/Team:TJUSLS_China/Attributions">ATTRIBUTIONS</a>
−
                        </li>
+
  
−
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Human_Practices">HUMAN PRACTICES</a>
+
                </li>
−
                            <ul>
+
−
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/HP/Silver">Silver</a></li>
+
−
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/HP/Gold">Gold</a></li>
+
−
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Integrated_Practices">Integrated Practices</a></li>
+
−
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Engagement">Engagement</a></li>
+
−
                             
+
  
−
                            </ul>
+
                <li onmouseover="getHover(5)" class=""><a href="https://2016.igem.org/Team:TJUSLS_China/Human_Practices">HUMAN PRACTICES</a>
−
</li>
+
                    <ul>
−
                          
+
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/HP/Silver">Silver</a></li>
−
                         <li><a href="https://2016.igem.org/Team:TJUSLS_China/Team">TEAM</a>
+
                         <li><a href="https://2016.igem.org/Team:TJUSLS_China/HP/Gold">Gold</a></li>
 +
                         <li><a href="https://2016.igem.org/Team:TJUSLS_China/Integrated_Practices">Integrated Practices</a></li>
 +
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Engagement">Engagement</a></li>
 +
                    </ul>
 +
                </li>
 +
                <li onmouseover="getHover(6)" class=""><a href="https://2016.igem.org/Team:TJUSLS_China/Modeling">MODELING</a>
 +
                    <ul>
 +
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Modeling">Modeling</a></li>
 +
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Software">Software</a></li>
 +
                    </ul>
 +
                </li>
 +
                <li onmouseover="getHover(7)" class=""><a href="https://2016.igem.org/Team:TJUSLS_China/Team">TEAM</a>
  
−
                            <ul>
+
                    <ul>
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        <ul>
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            <li><a href="#part1">Stucture</a></li>
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            <li><a href="#part2">Surface display in E.coli</a></li>
  
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    Results
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        <h3 id="part1">Training Protein</h3>
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        <div class="sec-wenzi-content">This year, we hold couples of forums in order to talk with great biology professors about our project. According to the ardent brainstorm, we proposed a advanced theory and built a proof of concept system called Training Protein System. All the work we did was centered on validating whether the system is applicable to all enzymes or not.
 +
             Training Protein System is a universalizable system for all the protein enzymes. Its main idea is a kind of research ideas, which is the most unique thing of it. Specifically, first and the foremost, researchers should analyze the protein tertiary structure and find its enzyme active center. The next step has two paths to improve the activity of enzymes. One was to mutate its gene sequence; the other was to make a surface display separately using E.coli and Yeast.
 +
            We demonstrate it working under real condition. This year, we use the system on PETase to ensure if this system can apply in real practice. What is particularly noteworthy is we used the hydrophobic protein to modify the surface of reaction system to increase its activity. It is really lucky that we managed to have a satisfactory result in training PETase to work better.
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        </div>
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        <h3 id="part2">The degradation of normal plastics</h3>
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        <div class="sec-wenzi-content">To find a better way to solve the problem of white pollution, we went to PET factory for a visit to learn more about PET and its industry. In factory, we know the latest technology of PET. Before we leaving, we get some PET samples from the director of the factory. It really interest us result in that we only use the standardized PET samples which are only used in labs for degradation and HPLC detection. This time we can do that on the normal plastic which we called hcPET in order to show our system working under real word conditions. Our HPLC results is shown as following.</div>
  
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        <div class="sec-wenzi-content">Figure 1. Relative enzyme activity of  engineering bacteria E.coli(BL21)/pET22b(+)LAP when they degarade PET bottles in different contitions.</div>
  
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        <h3 id="part3">Exploring for the best conditions by modeling</h3>
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        <div class="sec-wenzi-content">In order to apply the tech in the project into reality, we design a software which can assist us in processing data of HPLC. Our software can calculate peak area of any substance by the raw data of HPLC conveniently and accurately. It can display the image details saved at any period of time. It also can automatically identify substance peak and calculate the area. In addition, we can manually select the range of the area calculated. What’s more, in order to facilitate the analysis, it can store the results of calculations in a same file which contains the filename of the HPLC data.
 +
            Our software can be used on Windows system. The installation manual is attached to our software. And we have a user guide to explain how to operate our software conveniently.
 +
            Meanwhile, we built a multivariate nonlinear regression analysis model to predict the optimum condition of recombination cell decomposing PET. We set three variables as the reaction conditions in our model, which respectively were the cell concentration, the time and temperature of protein-induced expression. Then we established a multivariate nonlinear regression equation after the reasonable analysis.
 +
            Finally, we calculated the optimal condition of PET decomposition which means we can make the degradation of the PET much more efficient.
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        </div>
  
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                        <h3>welcome!</h3>
 
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                        <p>Quis autem vel eum iure reprehenderit, qui in ea voluptate velit esse, quam nihil molestiae consequatur, vel illum, qui dolorem eum fugiat, quo voluptas nulla pariatur. Sed in lacus ut enim adipiscing aliquet. Nulla venena tis. In pede mi, aliquet sit amet, euismod in, auctor ut, ligula. Aliquam dapibus tincidunt metus. Praesent justo dolor, lobortis quis, lobortis dignissim, pulvinar ac, lorem. Vestibulum sed ante. Donec sagittis euismod purus.Dolor nunc vule putateulr ips dol consec.Donec semp ertet laciniate ultricie upien disse comete dolo lectus fgilla itollicil tua ludin dolor nec met quam accumsan dolore. </p>
 
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                    <h3>testimonial</h3>
 
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                <div class="test-monials">
 
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                    &lt;!&ndash; start content_slider &ndash;&gt;
 
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                                <h4><a href="#">John Doe</a></h4>
 
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                                <h4><a href="#">Mark Trand</a></h4>
 
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        <!---- testmonials ---->
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        <div class="copyrights">Collect from <a href="http://www.cssmoban.com/" >aaaa</a></div>
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                <h3>Recent from the event</h3>
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                        <p class="link">09/05/2015 by<a href="#">Admin</a>13 comments</p>
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                        <h4><a href="#">Vestibulum iaculis</a></h4>
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                        <p>Raemuis ultricaretra gc accumsan malda or sit amet errepsm do elentu vel curslor sitmt conaiing.</p>
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                        <p class="link">19/8/2013 by<a href="#">Admin</a>10 comments</p>
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                        <h4><a href="#">Pellentesque sed dolor</a></h4>
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                        <p>Raemuis ultricaretra gc accumsan malda or sit amet errepsm do elentu vel curslor sitmt conaiing.</p>
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                        <p class="link">29/9/2015 by<a href="#">Admin</a>09 comments</p>
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                        <h4><a hrf="#">Vestibulum iaculis</a></h4>
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                        <p>Raemuis ultricaretra gc accumsan malda or sit amet errepsm do elentu vel curslor sitmt conaiing.</p>
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                    <div style="background-color: #eee">
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                        <div id="part1">
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                            <p>
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                                1111111111111111111111111111111111111111111111111
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                                PETase
+
−
                                PET降解酶发现于一种以PET为主要碳源的微生物（Ideonella sakaiensis 201-F6），该酶能将大分子聚合物降解为单体。表面展示技术是通过目的蛋白或多肽的编码基因与锚定蛋白的基因融合，通过融合蛋白的后翻译、折叠，将目的蛋白展示在宿主细胞壁的表面，从而获得全细胞催化剂的一种方法。我团队课题以PETase为主体，利用表面展示技术，从三个方面进行研究：
+
−
                                第一，通过蛋白质晶体学以及X射线衍射技术来解析PETase的结构，找到酶的催化中心和结合中心，并依据其结构特性指导突变位点的选择，从而对PETase进行定向突变以提高其降解效率和热稳定性；第二，将其进行原核（大肠杆菌）和真核（毕赤酵母）表面展示以进行全细胞酶催化反应。第三，将其与疏水蛋白在毕赤酵母中进行融合分泌，利用疏水蛋白两面性中的强疏水性为PET降解反应提供疏水环境以促进降解效率。同时，将疏水蛋白与PETase在毕赤酵母中进行共展示，利用疏水蛋白能改造细胞表面性质以适应极端环境的特点进一步提高全细胞催化剂的降解效率，打破了反应条件的限制，极大地拓宽了PETase降解反应的条件，以实现PETase的工业应用。
+
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                            </p>
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                        <div id="part2">
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                            <p>
+
−
                                2222222222222222222222222222222222222222222222222
+
−
                                PETase
+
−
                                PET降解酶发现于一种以PET为主要碳源的微生物（Ideonella sakaiensis 201-F6），该酶能将大分子聚合物降解为单体。表面展示技术是通过目的蛋白或多肽的编码基因与锚定蛋白的基因融合，通过融合蛋白的后翻译、折叠，将目的蛋白展示在宿主细胞壁的表面，从而获得全细胞催化剂的一种方法。我团队课题以PETase为主体，利用表面展示技术，从三个方面进行研究：
+
−
                                第一，通过蛋白质晶体学以及X射线衍射技术来解析PETase的结构，找到酶的催化中心和结合中心，并依据其结构特性指导突变位点的选择，从而对PETase进行定向突变以提高其降解效率和热稳定性；第二，将其进行原核（大肠杆菌）和真核（毕赤酵母）表面展示以进行全细胞酶催化反应。第三，将其与疏水蛋白在毕赤酵母中进行融合分泌，利用疏水蛋白两面性中的强疏水性为PET降解反应提供疏水环境以促进降解效率。同时，将疏水蛋白与PETase在毕赤酵母中进行共展示，利用疏水蛋白能改造细胞表面性质以适应极端环境的特点进一步提高全细胞催化剂的降解效率，打破了反应条件的限制，极大地拓宽了PETase降解反应的条件，以实现PETase的工业应用。
+
−
                            </p>
+
−
                            <img src="https://static.igem.org/mediawiki/2016/7/77/B1.jpg" alt="/">
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−
                        </div>
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                        <div id="part3">
+
−
                            <p>
+
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                                333333333333333333333333333333333333333333333333
+
−
                                PETase
+
−
                                PET降解酶发现于一种以PET为主要碳源的微生物（Ideonella sakaiensis 201-F6），该酶能将大分子聚合物降解为单体。表面展示技术是通过目的蛋白或多肽的编码基因与锚定蛋白的基因融合，通过融合蛋白的后翻译、折叠，将目的蛋白展示在宿主细胞壁的表面，从而获得全细胞催化剂的一种方法。我团队课题以PETase为主体，利用表面展示技术，从三个方面进行研究：
+
−
                                第一，通过蛋白质晶体学以及X射线衍射技术来解析PETase的结构，找到酶的催化中心和结合中心，并依据其结构特性指导突变位点的选择，从而对PETase进行定向突变以提高其降解效率和热稳定性；第二，将其进行原核（大肠杆菌）和真核（毕赤酵母）表面展示以进行全细胞酶催化反应。第三，将其与疏水蛋白在毕赤酵母中进行融合分泌，利用疏水蛋白两面性中的强疏水性为PET降解反应提供疏水环境以促进降解效率。同时，将疏水蛋白与PETase在毕赤酵母中进行共展示，利用疏水蛋白能改造细胞表面性质以适应极端环境的特点进一步提高全细胞催化剂的降解效率，打破了反应条件的限制，极大地拓宽了PETase降解反应的条件，以实现PETase的工业应用。
+
−
                            </p>
+
−
                            <img src="https://static.igem.org/mediawiki/2016/7/77/B1.jpg" alt="/">
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                        </div>
+
−
                        <div id="part4">
+
−
                            <p>
+
−
                                44444444444444444444444444444444444444444444444444
+
−
                                PETase
+
−
                                PET降解酶发现于一种以PET为主要碳源的微生物（Ideonella sakaiensis 201-F6），该酶能将大分子聚合物降解为单体。表面展示技术是通过目的蛋白或多肽的编码基因与锚定蛋白的基因融合，通过融合蛋白的后翻译、折叠，将目的蛋白展示在宿主细胞壁的表面，从而获得全细胞催化剂的一种方法。我团队课题以PETase为主体，利用表面展示技术，从三个方面进行研究：
+
−
                                第一，通过蛋白质晶体学以及X射线衍射技术来解析PETase的结构，找到酶的催化中心和结合中心，并依据其结构特性指导突变位点的选择，从而对PETase进行定向突变以提高其降解效率和热稳定性；第二，将其进行原核（大肠杆菌）和真核（毕赤酵母）表面展示以进行全细胞酶催化反应。第三，将其与疏水蛋白在毕赤酵母中进行融合分泌，利用疏水蛋白两面性中的强疏水性为PET降解反应提供疏水环境以促进降解效率。同时，将疏水蛋白与PETase在毕赤酵母中进行共展示，利用疏水蛋白能改造细胞表面性质以适应极端环境的特点进一步提高全细胞催化剂的降解效率，打破了反应条件的限制，极大地拓宽了PETase降解反应的条件，以实现PETase的工业应用。
+
−
                            </p>
+
−
                            <img src="https://static.igem.org/mediawiki/2016/7/77/B1.jpg" alt="/">
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                        </div>
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                        <div id="part5">
+
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                            <p>
+
−
                                55555555555555555555555555555555555555555555555555
+
−
                                PETase
+
−
                                PET降解酶发现于一种以PET为主要碳源的微生物（Ideonella sakaiensis 201-F6），该酶能将大分子聚合物降解为单体。表面展示技术是通过目的蛋白或多肽的编码基因与锚定蛋白的基因融合，通过融合蛋白的后翻译、折叠，将目的蛋白展示在宿主细胞壁的表面，从而获得全细胞催化剂的一种方法。我团队课题以PETase为主体，利用表面展示技术，从三个方面进行研究：
+
−
                                第一，通过蛋白质晶体学以及X射线衍射技术来解析PETase的结构，找到酶的催化中心和结合中心，并依据其结构特性指导突变位点的选择，从而对PETase进行定向突变以提高其降解效率和热稳定性；第二，将其进行原核（大肠杆菌）和真核（毕赤酵母）表面展示以进行全细胞酶催化反应。第三，将其与疏水蛋白在毕赤酵母中进行融合分泌，利用疏水蛋白两面性中的强疏水性为PET降解反应提供疏水环境以促进降解效率。同时，将疏水蛋白与PETase在毕赤酵母中进行共展示，利用疏水蛋白能改造细胞表面性质以适应极端环境的特点进一步提高全细胞催化剂的降解效率，打破了反应条件的限制，极大地拓宽了PETase降解反应的条件，以实现PETase的工业应用。
+
−
                            </p>
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Revision as of 15:43, 18 October 2016

Menu
Results

Training Protein

This year, we hold couples of forums in order to talk with great biology professors about our project. According to the ardent brainstorm, we proposed a advanced theory and built a proof of concept system called Training Protein System. All the work we did was centered on validating whether the system is applicable to all enzymes or not. Training Protein System is a universalizable system for all the protein enzymes. Its main idea is a kind of research ideas, which is the most unique thing of it. Specifically, first and the foremost, researchers should analyze the protein tertiary structure and find its enzyme active center. The next step has two paths to improve the activity of enzymes. One was to mutate its gene sequence; the other was to make a surface display separately using E.coli and Yeast. We demonstrate it working under real condition. This year, we use the system on PETase to ensure if this system can apply in real practice. What is particularly noteworthy is we used the hydrophobic protein to modify the surface of reaction system to increase its activity. It is really lucky that we managed to have a satisfactory result in training PETase to work better.

The degradation of normal plastics

To find a better way to solve the problem of white pollution, we went to PET factory for a visit to learn more about PET and its industry. In factory, we know the latest technology of PET. Before we leaving, we get some PET samples from the director of the factory. It really interest us result in that we only use the standardized PET samples which are only used in labs for degradation and HPLC detection. This time we can do that on the normal plastic which we called hcPET in order to show our system working under real word conditions. Our HPLC results is shown as following.
Figure 1. Relative enzyme activity of engineering bacteria E.coli(BL21)/pET22b(+)LAP when they degarade PET bottles in different contitions.

Exploring for the best conditions by modeling

In order to apply the tech in the project into reality, we design a software which can assist us in processing data of HPLC. Our software can calculate peak area of any substance by the raw data of HPLC conveniently and accurately. It can display the image details saved at any period of time. It also can automatically identify substance peak and calculate the area. In addition, we can manually select the range of the area calculated. What’s more, in order to facilitate the analysis, it can store the results of calculations in a same file which contains the filename of the HPLC data. Our software can be used on Windows system. The installation manual is attached to our software. And we have a user guide to explain how to operate our software conveniently. Meanwhile, we built a multivariate nonlinear regression analysis model to predict the optimum condition of recombination cell decomposing PET. We set three variables as the reaction conditions in our model, which respectively were the cell concentration, the time and temperature of protein-induced expression. Then we established a multivariate nonlinear regression equation after the reasonable analysis. Finally, we calculated the optimal condition of PET decomposition which means we can make the degradation of the PET much more efficient.

Copyright 2016 TJUSLS