Difference between revisions of "Team:SCUT-China/Members"

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{{SCUT-China/CSS-header}}
 +
{{SCUT-China/CSS-TTF}}
  
 +
 +
<html>
 +
<head>
 +
 +
  <meta charset="utf-8">
 +
  <title></title>
 +
  <!-- 4444444444444444444444此处上传时需要删除44444444444444444 -->
 +
  <link rel="stylesheet" href="css/style.css">
 +
  <link rel="stylesheet" href="css/style-content.css">
 +
  <link rel="stylesheet" href="css/style_ttc_p.css">
 +
  <!-- 4444444444444444444444此处上传时需要删除44444444444444444 -->
 +
 +
 +
 +
 +
      <style media="screen">
 +
        #top_title{ display:none; }
 +
        #content { padding:0px; width:100%; margin-top:-7px; margin-left:0px; border:none;}
 +
      body, html {background-color:#ececea; width: 100%; height: 100%;scroll-behavior: smooth;}
 +
 +
/* 666666666666666666666666666666666666666666666666666666666666以下为加载动画样式66666666666666666666666666666666666666666 */
 +
        #gif
 +
          {
 +
            position: fixed;
 +
            top: 0;
 +
            left: 0;
 +
            width: 100%;
 +
            height: 100%;
 +
            z-index: 99999;
 +
            /* display: -webkit-box; */
 +
            -webkit-box-orient: horizontal;
 +
            -webkit-box-pack: center;
 +
            -webkit-box-align: center;
 +
            background: #fff;
 +
        }
 +
 +
        #gif>img/*设置gif下的img元素样式*/
 +
        {
 +
 +
            /* width:40%; */
 +
        }
 +
          .gif2{
 +
            top: 25%;
 +
            height:auto;
 +
            margin-left:30%;
 +
            width:40%;
 +
          }
 +
          .gif1{
 +
            margin-left:35%;
 +
            top: 70%;
 +
            width:30%;
 +
            margin-top:-25px;
 +
 +
          }
 +
 +
/* 666666666666666666666666666666666666666666666666666666666666以上为加载动画样式66666666666666666666666666666666666666666 */
 +
  /* 左侧导航栏样式 */
 +
  #bodyContent h2{
 +
    padding:10px;
 +
  }
 +
  /* css */
 +
  html, body, div, span, applet, object, iframe,
 +
  h1, h2, h3, h4, h5, h6, p, blockquote, pre,
 +
  a, abbr, acronym, address, big, cite, code,
 +
  del, dfn, em, img, ins, kbd, q, s, samp,
 +
  small, strike, strong, sub, sup, tt, var,
 +
  b, u, i, center,
 +
  dl, dt, dd, ol, ul, li,
 +
  fieldset, form, label, legend,
 +
  table, caption, tbody, tfoot, thead, tr, th, td,
 +
  article, aside, canvas, details, embed,
 +
  figure, figcaption, footer, header, hgroup,
 +
  menu, nav, output, ruby, section, summary,
 +
  time, mark, audio, video {
 +
      margin: 0;
 +
      padding: 0;
 +
      border: 0;
 +
  }
 +
 +
  /* HTML5 display-role reset for older browsers */
 +
  article, aside, details, figcaption, figure,
 +
  footer, header, hgroup, menu, nav, section {
 +
      display: block;
 +
  }
 +
  body{
 +
    line-height: 1.5;
 +
  }
 +
  .middle{
 +
    background: #f4f3ee;
 +
    }
 +
  .middle_img{
 +
    width:100%;
 +
    height:100%;
 +
    background-color:rgba(0,0,0,0);
 +
    background:url("https://static.igem.org/mediawiki/2021/2/2f/T--SCUT-China--description-0.png") ;
 +
    background-size: 100% 100%;
 +
    background-attachment: fixed;
 +
    z-index: -1;
 +
    /* border-radius: 300px; */
 +
    /* background: #154889; */
 +
    overflow: hidden;
 +
  }
 +
 +
  ol, ul {
 +
      list-style: none;
 +
  }
 +
  blockquote, q {
 +
      quotes: none;
 +
  }
 +
  blockquote:before, blockquote:after,
 +
  q:before, q:after {
 +
      content: '';
 +
      content: none;
 +
  }
 +
  table {
 +
      border-collapse: collapse;
 +
      border-spacing: 0;
 +
  }
 +
  .left{
 +
    width: 25%;
 +
    height:auto;
 +
    float: left;
 +
    background:#ececea ;
 +
  }
 +
  .right{
 +
    width: 75%;
 +
    float: left;
 +
    z-index: 0;
 +
    background: #ececea;
 +
  }
 +
 +
  .siderBar{
 +
    /* 更改鼠标设置图片-------------------------------------------------------------------- */
 +
    /* cursor:url('img/youzi_jiantou-1.png'),auto; */
 +
    width: 95%;
 +
  }
 +
  #nav{
 +
    background: #ececea ;
 +
    top: 20px;
 +
    /* margin-top: 13%; */
 +
  }
 +
  #nav li{
 +
    margin-top: 10px;
 +
      width: 95%;
 +
      /* height: 50px; */
 +
      line-height: 25px;
 +
      text-align: center;
 +
      line-height:25px;
 +
      color: #2C2C2C;
 +
      margin-bottom: 10px;
 +
      padding-top:8px;
 +
      cursor: pointer;
 +
      font-family: 'H2_text';
 +
      font-weight:200;
 +
      font-size: 19px;
 +
      transition: all 0.2s ease;
 +
      -webkit-transition: all 0.2s ease;
 +
      -moz-transition: all 0.2s ease;
 +
  }
 +
  #nav li a{
 +
    padding-top: 15px;
 +
    color:#2C2C2C;
 +
    text-decoration:none;
 +
  }
 +
  #nav li a_nav{
 +
    margin-left: 20px;
 +
    padding-top: 20px;
 +
  }
 +
 +
  /* #nav li a:hover > img{
 +
    visibility:visible;
 +
  } */
 +
  #nav li.on{
 +
      background:#dad9d5;
 +
  }
 +
 +
 +
  #right-nav li a img{
 +
    width: 30px;
 +
    height: auto;
 +
    float: right;
 +
    margin-right: 10px;
 +
  }
 +
  #right-nav {
 +
    position: fixed;
 +
    right: 0px;
 +
    bottom: 50px;
 +
 +
  }
 +
  #right-nav li {
 +
        width: 167px;
 +
        height: 32px;
 +
        line-height: 32px;
 +
        text-align: center;
 +
        color: #ffffff;
 +
        margin-bottom: 5px;
 +
        background: url("images/side-nav-bg.jpg") no-repeat;
 +
        cursor: pointer;
 +
        transition: all 0.2s ease;
 +
        -webkit-transition: all 0.2s ease;
 +
        -moz-transition: all 0.2s ease;
 +
  }
 +
  #right-nav li.on{
 +
      transform: translate(-34px);
 +
      -webkit-transform: translate(-34px);
 +
      -moz-transform: translate(-34px);
 +
 +
  }
 +
  </style>
 +
 +
  <!-- 0000000000000000000000000000000000000右侧工具栏000000000000000000000000000000000 -->
 +
  <!-- 0000000000000000000000000000000000000右侧工具栏000000000000000000000000000000000 -->
 +
  <style>
 +
  .card-wrapper a:link,
 +
  .card-wrapper a:hover,
 +
  .card-wrapper a:visited,
 +
  .card-wrapper a:active {
 +
  color: #fff;
 +
  text-decoration: none;
 +
  }
 +
 +
  .card-holder {
 +
  position: fixed;
 +
  z-index: 9999;
 +
  width: 0px;
 +
  overflow: visible;
 +
  visibility: hidden;
 +
  }
 +
 +
  .card-wrapper {
 +
  display: inline-block;
 +
  float: left;
 +
  clear: both;
 +
  }
 +
 +
  .card {
 +
  position: relative;
 +
  right: 32px;
 +
  padding: 8px 32px 8px 32px;
 +
  margin: 8px;
 +
  x-box-shadow: 0 0 8px 0px rgba(0, 0, 0, 0.5);
 +
  box-shadow: 8px 0 8px -8px rgba(0, 0, 0, 0.5);
 +
  background: #fff;
 +
  transition: all 0.3s ease-in-out 0.1s;
 +
  }
 +
  .card_img {
 +
  position: relative;
 +
  right: 25px;
 +
  width:40px;
 +
  height:40px;
 +
  magin:10px;
 +
  bottom: -10px;
 +
  x-box-shadow: 0 0 8px 0px rgba(0, 0, 0, 0.5);
 +
  box-shadow: 8px 0 8px -8px rgba(0, 0, 0, 0.5);
 +
  background:rgba(0, 0, 0, 0);
 +
  }
 +
  .card-wrapper img{
 +
  margin-right: 32px;
 +
  width:100%;
 +
  height:auto;
 +
 +
  }
 +
 +
  .card:hover {
 +
  position: relative;
 +
  right: 100%;
 +
  margin-right: -32px;
 +
  box-shadow:
 +
  0 -8px 8px -8px rgba(0, 0, 0, 0.5),
 +
  0 8px 8px -8px rgba(0, 0, 0, 0.5);
 +
  transition: all 0.3s ease-in-out;
 +
  }
 +
 +
  .card-content {
 +
  display: inline-block;
 +
  color: #fff;
 +
  font-family: 'p_text';
 +
  font-size: 16px;
 +
  font-weight:450;
 +
  white-space: nowrap;
 +
  margin-right: 10px;
 +
  }
 +
 +
  .bg-01 { background: #539770; }
 +
  .bg-02 { background: #4B7D74; }
 +
  .bg-03 { background: #8DC2BC; }
 +
  .bg-04 { background: #EDD6B4; }
 +
  .bg-05 { background: #BE7467; }
 +
  .bg-06 { background: #E2AE63; }
 +
 +
  .card-right{
 +
  z-index: 999;
 +
  }
 +
  </style>
 +
  <!-- 0000000000000000000000000000000000000正文内容0000000000000000000000000000000 -->
 +
  <style>
 +
  .main_div{
 +
    background: #ececea;
 +
    display:inline-block;
 +
    width:100%;
 +
  }
 +
  #wrap{
 +
      width: 100%;
 +
  }
 +
  #wrap div{
 +
      /* width: 100%; */
 +
      /* height:auto; */
 +
      /* height: 500px; */
 +
  }
 +
  #content_part{
 +
    display:inline-block;
 +
    margin-top:10px;
 +
    margin-bottom: 15px;
 +
    /* background: #f4f3ee; */
 +
  }
 +
  #hrmar{
 +
      background: #f4f3ee;
 +
      height: auto;
 +
      margin-top:30px;
 +
      margin-right: 40px;
 +
      margin-bottom: 10px;
 +
      color:#333;
 +
 +
      filter:progid:DXImageTransform.Microsoft.Shadow(color=#909090,direction=120,strength=4);
 +
 +
      -moz-box-shadow: 2px 2px 10px #909090;
 +
 +
      -webkit-box-shadow: 2px 2px 10px #909090;
 +
 +
      box-shadow:2px 2px 10px #909090;
 +
  }
 +
  .content_div_img{
 +
    margin: 10px;
 +
    text-align: center;
 +
    justify-content: center;
 +
    display: flex;
 +
    /* width: 900px;
 +
    height: 500px; */
 +
 +
  }
 +
  .content_div_img img{
 +
    width:85%;
 +
    height:auto;
 +
  }
 +
 +
  .content_text_h2{
 +
    text-align: left;
 +
    margin-left: 70px;
 +
    margin-top: 10px;
 +
    font-size: 25px;
 +
    font-family:"H2_text";
 +
  }
 +
  .content_div_text{
 +
    letter-spacing: 0.4px;
 +
    margin: 20px;
 +
    padding-left:100px;
 +
    padding-right:100px;
 +
    /* text-align:center; */
 +
    float: left;
 +
    line-height: 1.5;
 +
    display: inline-block;
 +
    vertical-align: middle;
 +
    text-align: left;
 +
 +
  }
 +
  .content_div_text span{
 +
 +
  font-weight:bold;
 +
  }
 +
  .content_div_text p{
 +
    line-height: 1.5;
 +
    color: #2C2C2C;
 +
    text-align: justify;
 +
    font-family:"P_text";
 +
    word-wrap: break-word
 +
  }
 +
  hr{
 +
    margin-top: 1rem;
 +
    margin-bottom: 1rem;
 +
    border: 0;
 +
    border-top: 2px solid rgba(0,0,0,.1);
 +
    left: 20px;
 +
  }
 +
  .hrmar{
 +
    margin: 0rem 2rem 0 0;
 +
    width: 95%;
 +
  }
 +
  /* --------------------------------------------------------侧边进度条调整--------------------------------- */
 +
  .divBox_div{
 +
    position: fixed;
 +
    width: 10px;
 +
    height: 100%;
 +
    z-index: 99;
 +
  }
 +
  #divBox {
 +
      position: relative;
 +
      width: 10px;
 +
      background-color: #feb770;
 +
      padding: 1px;
 +
      text-align: center;
 +
      top:5%;}
 +
  #divBox_0 {
 +
          position: relative;
 +
          width:10px;
 +
          height: 100%;
 +
          background-color: #feebc3;
 +
          padding: 1px;
 +
          text-align: center;
 +
          top:5%;
 +
          visibility: hidden;
 +
        }
 +
  #bottom_div{
 +
    float: right;
 +
    width: 100%;
 +
    height: 35%;
 +
    /* background-color: #f4f3ee; */
 +
    background-color: #ececea;
 +
  }
 +
  @media only screen and (min-width: 1500px) and (max-width: 2560px) {
 +
    #bottom_div{
 +
      height: 33%;
 +
      height: 260px;
 +
 +
    }
 +
  }
 +
  @media only screen and (min-width: 1000px) and (max-width: 1500px) {
 +
    #bottom_div{
 +
      height: 30%;
 +
      height: 260px;
 +
    }
 +
  }
 +
  @media only screen and (min-width: 768px) and (max-width: 1199px) {
 +
    #bottom_div{
 +
      height: 220px;
 +
    }
 +
 +
  }
 +
  @media (max-width: 768px) {
 +
    #bottom_div{
 +
      height: 180px;
 +
    }
 +
  }
 +
  #bottom_div_img{
 +
    float: right;
 +
    width: 100%;
 +
    height: 100%;
 +
    margin-left: 30px;
 +
    background: url("https://static.igem.org/mediawiki/2021/6/67/T--SCUT-China--background-home-bottom.png");
 +
    background-size: 100% 100%;
 +
  }
 +
  #bottom_div .a_div{
 +
    display: relative;
 +
    width: 17%;
 +
    height:50px;
 +
    margin-left: 30px;
 +
    opacity: 0;
 +
  }
 +
  #bottom_div a{
 +
    opacity: 1;
 +
  }
 +
 +
  /* ----------------------------------------------------------------------------------轮播图 */
 +
  #parent{
 +
      position: relative;
 +
      margin: 10px auto;
 +
      padding: 0;
 +
      width: 500px;
 +
      height: 309px;
 +
  }
 +
 +
  #uls{
 +
      position: relative;
 +
      margin: 0;
 +
      padding: 0;
 +
      width: 500px;
 +
      height: 309px;
 +
      overflow: hidden;
 +
  }
 +
 +
  #img_ul{
 +
      position: absolute;
 +
      margin: 0;
 +
      padding: 0;
 +
      left: 0;
 +
      top: 0;
 +
      width: 3000px;          /*多留出一张图片的宽度！*/
 +
      list-style: none;
 +
  }
 +
  #img_ul li{
 +
      float: left;
 +
      margin: 0;
 +
      padding: 0;
 +
      width: 500px;
 +
      height: 309px;
 +
  }
 +
  #img_ul li img{
 +
      width: 500px;
 +
      height: 250px;
 +
  }
 +
 +
  #litCir_ul{
 +
      position: absolute;
 +
      margin: 0;
 +
      padding: 0;
 +
      right: 10px;
 +
      bottom: 10px;
 +
      list-style: none;
 +
  }
 +
  #litCir_ul li{
 +
      margin: 0;
 +
      padding: 0;
 +
      float: left;
 +
      width: 20px;
 +
      height: 20px;
 +
      text-align: center;
 +
      line-height: 20px;
 +
      border-radius: 50%;
 +
      margin-left:10px ;
 +
      cursor: pointer;
 +
  }
 +
 +
  #litCir_ul li.active{
 +
      background-color: white;
 +
  }
 +
  #litCir_ul li.quiet{
 +
      background-color: #feb770;
 +
  }
 +
 +
  #buttons{
 +
      margin: 0;
 +
      padding: 0;
 +
      display: none;
 +
  }
 +
  #buttons span{
 +
      position: absolute;
 +
      width: 40px;
 +
      height: 40px;
 +
      top: 50%;
 +
      margin-top: -20px;
 +
      line-height: 40px;
 +
      text-align: center;
 +
      font-weight: bold;
 +
      font-family: Simsun;
 +
      font-size: 30px;
 +
      border: 1px solid #fff;
 +
      opacity: 0.3;
 +
      cursor: pointer;
 +
      color: #fff;
 +
      background: black;
 +
  }
 +
  #left_button{
 +
      left: 5px;
 +
  }
 +
  #right_button{
 +
      left: 100%;
 +
      margin-left: -45px;
 +
  }
 +
  /* ------------------------------------------------------------------------------------------------------------------------------------------------------ */
 +
  </style>
 +
 +
      <title></title>
 +
    </head>
 +
 +
    <body  >
 +
 +
      <!-- 顶部导航栏位置 -->
 +
  <div class="header" >
 +
 +
        <!---Header Start-->
 +
      <div class="header-area">
 +
          <div class="main-header ">
 +
              <div class="header-bottom  header-sticky">
 +
                  <div class="container-fluid">
 +
                      <div class="menu-all">
 +
                          <!-- Logo -->
 +
                          <div class="logo">
 +
                              <a href="https://2021.igem.org/Team:SCUT-China"><img class="logoimg" src="https://static.igem.org/mediawiki/2021/5/5b/T--SCUT-China--logo.png" alt=""></a>
 +
                          </div>
 +
                          <!-- Main-menu -->
 +
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              <li style="font-size:26px;"><a class="a_nav">Description</a></li>
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              <li ><a>Overview</a></li>
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              <li><a >Inspiration </a></li>
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                <li><a >Nootka-Boom！</a></li>
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                    <h2  class="content_text_h2">Overview</h2>
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                        <p  id="p1_1"align="left">      A group of students from SCUT in Guangzhou came up with an idea to alleviate the scourge of mosquitoes. In inspiration, we skillfully used Baidu index big data analysis, investigated the official data of the WORLD Health Organization and EPA, and known a new mosquito repellent ingredient--Nootkatone. In the conception stage of our project, based on the rise of biological manufacturing and synthetic biology, we considered from three aspects: the selection of chassis cells, the identification of synthetic pathways and how to increase yield, so as to build a cell factory with high yield of Nootkatone and to realize our Nootka-Boom!                  </p>
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                        <p  id="p1_1"align="left">      A group of students from SCUT in Guangzhou came up with an idea to alleviate the scourge of mosquitoes. In inspiration, we skillfully used Baidu index big data analysis, investigated the official data of the WORLD Health Organization and EPA, and known a new mosquito repellent ingredient--Nootkatone. In the conception stage of our project, based on the rise of biological manufacturing and synthetic biology, we considered from three aspects: the selection of chassis cells, the identification of synthetic pathways and how to increase yield, so as to build a cell factory with high yield of Nootkatone and to realize our Nootka-Boom!                  </p>
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                      <h2  class="content_text_h2">Inspiration</h2>
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                      <h4 class="content_text_h2" style="font-size:18px;margin-top:20px;">Regional necessity:</h4>
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                      <p  id="p2_1"align="left">      South China University of Technology is located in the humid and rainy Guangzhou, where the warm climate with unclear seasons is not only suitable for human to live but also for mosquito breeding. Whether indoors or outdoors, the mosquito problem has always plagued us, as well as other people living in Guangzhou. According to the data of Baidu index, from 2013 to 2021, the users who search the keyword of mosquito repellent on the Internet are most from Guangdong. And in Guangdong, the most dominant users having searched relevant questions are from Guangzhou. This can show that the mosquito problem is a high concern and urgent problem for people around us.
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                      <p style="font-size:14px;text-align:center;color:#A5A8A6;">Image Source:baidu.com</p>
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                    <h4 class="content_text_h2" style="font-size:18px;margin-top:20px;">International necessity:</h4>
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                        <p  id="p2_2"align="left">      Mosquito bites bring not only itchy skin, but also the risk of contracting infections mosquito-borne diseases. Mosquitoes are vector organisms and diseases that use mosquitoes as vectors include yellow fever, dengue fever, epidemic B encephalitis, malaria, lymphatic filariasis, Chikungunya, West Nile virus disease, and Rift Valley fever. In the case of dengue fever, for example, WHO listed it as one of the top 10 threats to global health in 2019. About 390 million people in 128 countries are at risk of dengue fever, with an average of 96 million people infected each year, including asymptomatic carriers. The mortality rate has remained stable at around 0.12%, which means that thousands of people die each year from dengue infection.                    </p>
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                        <p style="font-size:14px;text-align:center;color:#A5A8A6;">Data Source: World Health Organization</p>
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                        <p  id="p2_3"align="left">      The most direct solution to the problem seems to be the use of insecticide. However, monitoring has found that insecticide resistance is widespread among malaria vectors. According to the World Health Organization, 65 of 89 malaria-endemic countries have reported at least one endemic vector resistant to pyrethroids since 2010. All four classes of insecticides currently used for adult malaria vector control, pyrethroids, organochlorines, carbamates and organophosphates, have spread widely in Africa, the Americas, Southeast Asia, the eastern Mediterranean and the Western Pacific. Moreover, malaria vectors in some parts of Africa are now able to survive in environments with high concentrations of insecticides, suggesting that resistance in some malaria vector organisms is now increasing over time.                    </p>
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                        <p style="font-size:14px;text-align:center;color:#A5A8A6;">Source: WHO Malaria Threats Map.</p>
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                        <p  id="p2_4"align="left">      In the EPA's report on new repellent ingredients, we noted that compared with existing repellent ingredients, Nootkatone can maintain high repellent effect stably for a long time. As proven by EPA's data, the high-purity Nootkatone not only has high safety, but also has no irritation, no skin sensitization, and is a qualified mosquito repellent ingredient. According to the EPA, the creation of a new effective insecticide ingredient,Nootkatone, will help address the growing insecticide resistance to other products currently in use. However, currently there is no popular product related to Nootkatone on the market because its current primary production method is plant-based extraction, which is expensive and difficult to produce in large quantities. In order to further enable the wider application of Nootkatone, to exploit its value and to explore more of its possibilities, we have undertaken our project - Nootka-Boom!
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                    <h2 class="content_text_h2">Nootka-Boom！</h2>
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                    <h4 class="content_text_h2" style="font-size:18px;margin-top:20px;">Source of project</h4>
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                      <p  id="p3_1_0"align="left">    Synthetic biology with engineering design concept, design and transformation of organism to have a goal, subvert the biological technique innovation, is expected to decode the human resources, environment and other fields important challenge to provide a new solution With the rapid development of synthetic biology, through the microbial production of high value-added compounds become attractive In April 2013,Nature published a study on artemisinin, a highly effective semi-synthetic antimalarial drug in <i>Saccharomyces cerevisiae</i>, which greatly promoted the industrialization of microbial semi-synthetic artemisinin. This is the most successful case of heterologous synthesis of high value-added products by microorganisms.
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                        <p  id="p3_1_1"align="left">    Nootkatone is a natural ingredient found in some Alaskan yellow cedar herbs and citrus fruits that is responsible for grapefruit's distinctive smell. Nootkatone has long been used in many food products that have a citrus flavor or smell. DVBD scientists have found that Nootkatone is an effective mosquito repellent and insecticide that can be used against mosquito ticks and other pests<sup>[1]</sup>.
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                        <p  id="p3_1_2"align="left">    There are three main production methods of Nootkatone: Plant extraction method, chemical synthesis and microbial synthesis. Terpenoids content in plants is usually very low, the extraction method is easy to cause serious damage to wild plant resources, and the planting and processing of its crops are affected by many factors such as climate, environment and transportation, so the yield, quality and price of Nootkatone have uncontrollable changes, and the economic feasibility is poor. At present, the industry is often through a precursor of the direct oxidation of price is relatively cheap Valencene to obtain Nootkatone. But the reaction usually involves some environmental unfriendly oxidant, such as heavy metal salts. By contrast, microbial synthesis method is not restricted by raw material Green clean production process, has a great advantage<sup>[2]</sup>. Such as the Nature The focus of the synthetic biology industry is rapidly shifting to the over $20 billion fine chemicals market, which will rapidly advance the heterologous biosynthesis of natural products Therefore, to meet the growing demand for Nootkatone, SCUT-China wants to use synthetic biology methods to create a cell factory with high-yield of Nootkatone.
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                      <h4 class="content_text_h2" style="font-size:18px;margin-top:20px;">Selection of chassis cell</h4>
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                      <p  id="p3_2"align="left">    In the field of biomanufacturing, the selection of different chassis cells plays a key role in the biosynthesis of terpenoids. IPP and DMAPP, the precursors of terpenoids in E. coli, are mainly used for prenylation of tRNA and synthesis of FPP, and then for synthesis of quinones and cell walls[3]. Therefore, these chassis cells are mainly used for biosynthesis of alcohols and acids rather than terpenoids. In addition, due to the lack of post-translational modification, it is difficult for bacteria to express cytochrome P450, and many terpenoids have antibacterial activity. However, <i>Saccharomyces cerevisiae</i> has been widely used in terpenoid biosynthesis because it is generally regarded as safe (GRAS),10 industrially robust and able to functionally express eukaryotic cytochrome P450 enzymes[4]. Therefore, SCUT-China will also consider using <i>S. cerevisiae</i> to build our Nootkatone cell factory.
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                      <h4 class="content_text_h2" style="font-size:18px;margin-top:20px;">Identification of biosynthetic pathways</h4>
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                      <p  id="p3_3"align="left">          All terpenes are biosynthesized through mevalonate pathway (MVA) in yeast, originating from acetyl-CoA. And the intermediate product farnesyl diphosphate (FPP) is the direct precursor of valencene, which is catalyzed by valencene synthase (VS) to synthesize valencene. Then, valencene is oxidated to β-Nootkatol and (+)-nootkatone by HPO, AtCPR and ADH[2]. SCUT-China will introduce the Nootkatone synthesis pathway into <i>S. cerevisiae</i> to build our cell factory.
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                      <h4 class="content_text_h2" style="font-size:18px;margin-top:20px;">Idea to realize Nootka-Boom!</h4>
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                      <p  id="p3_4"align="left">    After constructing a <i>saccharomyces cerevisiae</i> cell factory capable of producing Nootkatone, we found that the construction of metabolic pathways usually involves the expression of multiple genes, whose expression levels span several orders of magnitude, and the regulation of gene expression by endogenous gene regulatory elements alone is not sufficient[5]. In addition, we also found that CnVS is the limiting enzyme in the metabolic pathway of Nootkatone synthesis, and its expression level plays a key role in Nootkatone production[2]. How to effectively improve the expression level of VS to improve Nootkatone precursor (valencene) and Nootkatone production is a big problem we are facing.<br><br>To solve the above two problems, we turned our attention to promoters that control gene transcription[6]. As the initiation of gene transcription, promoters are key components of cell factory design and metabolic pathway modification. Due to the limited number of well-characterized promoters in yeast and their small dynamic ranges, it is often difficult to satisfy the fine regulation of genes, optimize metabolic flux, and improve the yield of the target product[7]. Therefore, SCUT-China wants to obtain high-strength promoters to increase the expression of CnVS through engineering modification of <i>S. cerevisiae</i> natural promoters, and to obtain a good dynamic promoter library, which will provide useful and efficient help for the improvement of metabolic flux optimization target product yield and the design of cell factory.<br><br>In labs, our promoter engineering will be carried out from <a href="https://2021.igem.org/Team:SCUT-China/Design">Wet Lab </a>and <a href="https://2021.igem.org/Team:SCUT-China/Model">Dry Lab</a> respectively. Wet Lab mainly focuses on UAS combining different promoters to construct a series of hybrid-promoters with different strengths. Dry Lab obtained promoters with different intensities mainly from nucleosome affinity of promoters.
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                    <h2  class="content_text_h2">References</h2>
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                      <p>1. Clarkson, T.C., et al., Nootkatone Is an Effective Repellent against Aedes aegypti and Aedes albopictus. INSECTS, 2021. 12(5).    </p>
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                      <p>2. Ouyang, X., et al., Stepwise engineering of Saccharomyces cerevisiae to produce (+)-valencene and its related sesquiterpenes. RSC ADVANCES, 2019. 9(52): p. 30171-30181.  </p>
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                      <p>3. Wang, D., Z. Dai, and X. Zhang, [Production of plant-derived natural products in yeast cells - A review]. Wei sheng wu xue bao = Acta microbiologica Sinica, 2016. 56(3): p. 516-29.    </p>
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                      <p>4. Dai, Z.B., et al., Yeast synthetic biology for high-value metabolites. Fems Yeast Research, 2015. 15(1): p. 11.    </p>
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                      <p>5. Chen, X., et al., DCEO Biotechnology: Tools To Design, Construct, Evaluate, and Optimize the Metabolic Pathway for Biosynthesis of Chemicals. Chemical Reviews, 2018. 118(1): p. 4-72.    </p>
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                      <p>6. Young, E. and H. Alper, Synthetic Biology: Tools to Design, Build, and Optimize Cellular Processes. Journal of Biomedicine and Biotechnology, 2010.    </p>
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                      <p>7. Maury, J., et al., Reconstruction of a bacterial isoprenoid biosynthetic pathway in Saccharomyces cerevisiae. Febs Letters, 2008. 582(29): p. 4032-4038    </p>
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                    //- ------------------------------------以下为一个单位的替换需求，【注意id与之前对应段落相同】【注意两两对应，与以下对应】--------------------------------------------------------------------------------------
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                    document.getElementById("p1_1").innerHTML="      一群来自广州的华南理工大学学子萌发了要缓解蚊子困扰的想法。头脑风暴阶段，我们巧妙地使用到了百度指数大数据分析，调研了世界卫生组织、EPA等的官方数据，同时惊喜地发现了新型驱蚊成分——Nootkatone，在项目构思阶段，基于生物制造与合成生物学的兴起，我们从底盘细胞的选择、合成途径的构建、实现高产的想法三个方面考虑，打造能够高产圆柚酮的细胞工厂，实现Nootka-Boom!";
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                  document.getElementById("p2_1").innerHTML="      华南理工大学坐落于潮湿多雨的广州，四季不分明的温暖气候不仅适于人类居住，也适合于蚊虫孳生。无论是在室内还是室外，蚊虫问题一直困扰着我们，也困扰着生活在广州的其他人们。根据百度指数的数据显示，从2013年到2021年，在网络上检索关键词为驱蚊的用户最主要来自广东。而在广东中，最主要的检索用户来自广州。这可以说明，驱蚊问题对于广州人民来说是一个关注度很高，也亟待解决的问题。";
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                    document.getElementById("p2_2").innerHTML="      蚊虫叮咬带来的不仅是皮肤瘙痒，还有感染蚊媒传染病的风险。蚊子是一种病媒生物，以蚊子作为传播媒介的疾病包括黄热病、登革热、流行性乙型脑炎、疟疾、淋巴丝虫病、Chikungunya、西尼罗病毒病和裂谷热等。以登革热为例，世卫组织将登革热列为2019年全球健康面临的10大威胁之一。在128个国家中，约有3.9亿人面临登革热的风险，每年平均有9600万人感染，包括无症状携带者。死亡率稳定地维持在0.12%左右，这表示每年都有数千人因感染登革热死亡。";
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                    document.getElementById("p2_3").innerHTML="      面对病媒生物蚊子，最直接的解决方法就是使用蚊子的杀灭剂。然而监测发现，杀虫剂抗药性在疟疾病媒中广泛存在。据世界卫生组织的数据，自2010年以来，89个疟疾流行国家中有65个报告，至少有一个地方病媒对拟除虫菊酯产生抗药性。目前用于成人疟疾病媒控制的所有四类杀虫剂，拟除虫菊酯、有机氯、氨基甲酸盐和有机磷酸酯已经在非洲、美洲、东南亚、地中海东部和西太平洋地区广泛蔓延。而且非洲某些地区的疟疾病媒现在能够在高浓度杀虫剂的环境中存活，这表明目前某些疟疾病媒生物的抗药性随着时间的变迁正在逐渐增。";
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                    document.getElementById("p2_4").innerHTML="      在EPA关于新驱蚊成分的报道中，我们注意到了和现有驱蚊成分相比，能长期稳定保持高驱蚊效果的圆柚酮。由EPA的数据证明，高纯度的圆柚酮不仅具有高安全性，还没有刺激性、皮肤致敏性，是一种合格的驱蚊成分。而且根据EPA的说法，新的有效的杀虫剂成分——圆柚酮的产生，将有助于解决对目前使用的其他产品日益增长的杀虫剂耐药性。然而目前市面上圆柚酮相关产品并没有流行，因为目前其主要生产方式为植物提取，价格昂贵，难以大批量生产。为了进一步让圆柚酮能够被更广泛地应用，发挥其价值，挖掘其更多的可能性，我们开展了我们的项目——Nootka-Boom！";
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                    document.getElementById("p3_1_0").innerHTML="    合成生物学以工程化设计理念，对生物体进行有目标的设计与改造，形成生物技术颠覆式创新，有望为破解人类面临的资源、环境等领域重大挑战提供新的解决方案。随着合成生物学的快速发展，通过微生物改造生产高附加值化合物变得极具吸引力。2013年4月，Nature发表了在酿酒酵母中高效半合成抗疟疾药物青蒿素的研究，将微生物半合成青蒿素的产业化进程大步向前推进。这是微生物异源合成高附加值产品的最成功案例。";
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                    document.getElementById("p3_1_1").innerHTML="    Nootkatone是一种存在于阿拉斯加黄杉、一些草药和柑橘类水果中的天然成分，它是导致葡萄柚独特气味的原因。长期以来，诺卡通一直被用于许多具有柑橘风味或气味的食品产品中。DVBD的科学家发现诺卡酮是一种有效的驱蚊剂和杀虫剂，可以用来对付蚊子、扁虱和其他害虫。";
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                    document.getElementById("p3_1_2").innerHTML="    圆柚酮的生产方式主要有3种: 植物提取法、化学合成法和微生物合成法。萜类化合物在植物中的含量通常都很低，植物提取法对野生植物资源易造成严重破坏，且其作物的种植加工受到气候、环境、运输等诸多因素的影响，使得圆柚酮香料在产量、质量、价格方面存在不可控的变化，经济可行性较差。目前，工业上常常通过直接氧化价格相对便宜的前体物质瓦伦西亚烯来获得圆柚酮。但是，该反应过程通常涉及一些非环境友好的氧化剂，如三氧化铬等重金属盐类，相比之下，微生物合成方法不受原料的限制、生产过程绿色清洁，具有很大的优势。如Nature Reviews Microbiology文章所述：合成生物行业关注焦点正快速转变到价值超过200亿美元的“精细化学品”市场，其将快速推动天然产物的异源生物合成。因此，为了满足圆柚酮愈发增长的需求，SCUT-China希望利用合成生物学的手段来打造高产圆柚酮的细胞工厂。";
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                    document.getElementById("p3_2").innerHTML="      在生物制造这一领域中，选择不同的底盘细胞对于萜类化合物的生物合成有着关键性的影响。大肠杆菌中萜类前体IPP和DMAPP主要用于tRNA的异戊烯化和FPP的合成，进而用于醌类和细胞壁的合成，因此这类底盘细胞主要用于醇、酸等化学品而非萜类的生物合成。此外，由于细菌缺少翻译后修饰，难以表达细胞色素P450，且很多萜类化合物具有抗菌活性。因此，SCUT-China也将考虑使用酿酒酵母来打造我们的圆柚酮细胞工厂。 ";
 +
                    document.getElementById("p3_3").innerHTML="      在酵母中所有萜类化合物都是由乙酰辅酶A通过甲羟戊酸途径(MVA)合成的。中间体法尼基二磷酸(FPP)是瓦伦西亚烯的直接前体，在瓦伦西亚烯合成酶(VS)的催化下合成瓦伦西亚烯。然后，通过HPO、AtCPR和ADH[2]酶依次被氧化成β-Nootkatol和(+)nootkatone。因此，SCUT-China也将计划将圆柚酮合成途径引入酿酒酵母，以此建立我们的细胞工厂。";
 +
                    document.getElementById("p3_4").innerHTML="      在构建好能够生产圆柚酮的酿酒酵母细胞工厂后，我们发现代谢途径的构建通常涉及多个基因的表达，这些基因的表达水平横跨几个数量级，仅仅使用内源性的基因调控元件进行基因表达的调控是远远不够的，代谢途径的紊乱不仅会增加细胞的负担，导致细胞生长缓慢，还会造成生产目标产物不稳定性等。另外，我们还发现在合成圆柚酮的代谢途径中，CnVS是其中的限速酶，其表达水平对于圆柚酮产量具有关键作用，如何有效地提高VS的表达水平以提高圆柚酮前体（瓦伦西亚烯）以及圆柚酮的产量是我们面临的一大问题。<br><br>针对以上两个问题，我们将目光投向了控制基因转录的——启动子。作为基因转录的起始，启动子在细胞工厂设计和代谢途径改造中发挥重要的作用（Promoters are key components of cell factorydesign, allowing precise expression of genes in a heterologous pathway.）。由于酵母中表征良好的启动子数量有限，且动态范围小，通常很难满足对基因的精细调控和优化代谢通量，也很难实现目标产物产量的提高。酿酒酵母内源性启动子分为组成型和诱导型。诱导型启动子是可以在诱导剂存在下或在某些培养条件下启动表达的启动子，但添加外源诱导物会增加成本，并且有些诱导物对细胞有毒害。组成型启动子的特点是表达稳定，可以用于代谢途径的组装与优化，但只能维持相对稳定的转录水平，其动态范围差，阻碍了在代谢工程改造中的应用。<br><br>因此SCUT-China希望通过对酿酒酵母天然启动子进行工程改造，不仅获得高强度的启动子来增加VS的表达量，而且得到动态良好的启动子库，为代谢通量优化、目标产物产量的提升以及最终细胞工厂的设计提供有用且高效的帮助。<br><br>在lab 活动中，我们将分别从Wet Lab and Dry Lab两个方向进行具体的启动子工程改造，wet lab主要聚焦于组合不同启动子的UAS来构建一系列不同强度的杂合启动子；而dry lab主要从启动子与核小体的亲和力的角度考虑得到强度不同的启动子。";
 +
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                    // -------------------------- ------------------------------------以下为一个单位的替换需求，注意id与之前对应段落相同，【注意两两对应，与以上对应】--------------------------------------------------------------------------------------
 +
                    document.getElementById("p1_1").innerHTML="    A group of students from SCUT in Guangzhou came up with an idea to alleviate the scourge of mosquitoes. In inspiration, we skillfully used Baidu index big data analysis, investigated the official data of the WORLD Health Organization and EPA, and known a new mosquito repellent ingredient--Nootkatone. In the conception stage of our project, based on the rise of biological manufacturing and synthetic biology, we considered from three aspects: the selection of chassis cells, the identification of synthetic pathways and how to increase yield, so as to build a cell factory with high yield of Nootkatone and to realize our Nootka-Boom!";
 +
                    // -------------------------------------------------------以上为一个单位的替换代码-----------------------------------------------------------------------------------------------------------------
 +
                    document.getElementById("p2_1").innerHTML="      South China University of Technology is located in the humid and rainy Guangzhou, where the warm climate with unclear seasons is not only suitable for human to live but also for mosquito breeding. Whether indoors or outdoors, the mosquito problem has always plagued us, as well as other people living in Guangzhou. According to the data of Baidu index, from 2013 to 2021, the users who search the keyword of mosquito repellent on the Internet are most from Guangdong. And in Guangdong, the most dominant users having searched relevant questions are from Guangzhou. This can show that the mosquito problem is a high concern and urgent problem for people around us.  ";
 +
                    document.getElementById("p2_2").innerHTML="      Mosquito bites bring not only itchy skin, but also the risk of contracting infections mosquito-borne diseases. Mosquitoes are vector organisms and diseases that use mosquitoes as vectors include yellow fever, dengue fever, epidemic B encephalitis, malaria, lymphatic filariasis, Chikungunya, West Nile virus disease, and Rift Valley fever. In the case of dengue fever, for example, WHO listed it as one of the top 10 threats to global health in 2019. About 390 million people in 128 countries are at risk of dengue fever, with an average of 96 million people infected each year, including asymptomatic carriers. The mortality rate has remained stable at around 0.12%, which means that thousands of people die each year from dengue infection.";
 +
                    document.getElementById("p2_3").innerHTML="      The most direct solution to the problem seems to be the use of insecticide. However, monitoring has found that insecticide resistance is widespread among malaria vectors. According to the World Health Organization, 65 of 89 malaria-endemic countries have reported at least one endemic vector resistant to pyrethroids since 2010. All four classes of insecticides currently used for adult malaria vector control, pyrethroids, organochlorines, carbamates and organophosphates, have spread widely in Africa, the Americas, Southeast Asia, the eastern Mediterranean and the Western Pacific. Moreover, malaria vectors in some parts of Africa are now able to survive in environments with high concentrations of insecticides, suggesting that resistance in some malaria vector organisms is now increasing over time.";
 +
                    document.getElementById("p2_4").innerHTML="      In the EPA's report on new repellent ingredients, we noted that compared with existing repellent ingredients, Nootkatone can maintain high repellent effect stably for a long time. As proven by EPA's data, the high-purity Nootkatone not only has high safety, but also has no irritation, no skin sensitization, and is a qualified mosquito repellent ingredient. According to the EPA, the creation of a new effective insecticide ingredient, Nootkatone, will help address the growing insecticide resistance to other products currently in use. However, currently there is no popular product related to Nootkatone on the market because its current primary production method is plant-based extraction, which is expensive and difficult to produce in large quantities. In order to further enable the wider application of Nootkatone, to exploit its value and to explore more of its possibilities, we have undertaken our project - Nootka-Boom!";
 +
 +
                    document.getElementById("p3_1_0").innerHTML="    Synthetic biology with engineering design concept, design and transformation of organism to have a goal, subvert the biological technique innovation, is expected to decode the human resources, environment and other fields important challenge to provide a new solution With the rapid development of synthetic biology, through the microbial production of high value-added compounds become attractive In April 2013,Nature published a study on artemisinin, a highly effective semi-synthetic antimalarial drug in <i>Saccharomyces cerevisiae</i>, which greatly promoted the industrialization of microbial semi-synthetic artemisinin. This is the most successful case of heterologous synthesis of high value-added products by microorganisms.";
 +
                    document.getElementById("p3_1_1").innerHTML="    Nootkatone is a natural ingredient found in some Alaskan yellow cedar herbs and citrus fruits that is responsible for grapefruit's distinctive smell. Nootkatone has long been used in many food products that have a citrus flavor or smell. DVBD scientists have found that Nootkatone is an effective mosquito repellent and insecticide that can be used against mosquito ticks and other pests<sup>[1]</sup>.";
 +
                    document.getElementById("p3_1_2").innerHTML="    There are three main production methods of Nootkatone: Plant extraction method, chemical synthesis and microbial synthesis. Terpenoids content in plants is usually very low, the extraction method is easy to cause serious damage to wild plant resources, and the planting and processing of its crops are affected by many factors such as climate, environment and transportation, so the yield, quality and price of Nootkatone have uncontrollable changes, and the economic feasibility is poor. At present, the industry is often through a precursor of the direct oxidation of price is relatively cheap Valencene to obtain Nootkatone. But the reaction usually involves some environmental unfriendly oxidant, such as heavy metal salts. By contrast, microbial synthesis method is not restricted by raw material Green clean production process, has a great advantage<sup>[2]</sup>. Such as the Nature The focus of the synthetic biology industry is rapidly shifting to the over $20 billion fine chemicals market, which will rapidly advance the heterologous biosynthesis of natural products Therefore, to meet the growing demand for Nootkatone, SCUT-China wants to use synthetic biology methods to create a cell factory with high-yield of Nootkatone.";
 +
                    document.getElementById("p3_3").innerHTML="    All terpenes are biosynthesized through mevalonate pathway (MVA) in yeast, originating from acetyl-CoA. And the intermediate product farnesyl diphosphate (FPP) is the direct precursor of valencene, which is catalyzed by valencene synthase (VS) to synthesize valencene. Then, valencene is oxidated to β-Nootkatol and (+)-nootkatone by HPO, AtCPR and ADH[2]. SCUT-China will introduce the Nootkatone synthesis pathway into <i>S. cerevisiae</i> to build our cell factory.";
 +
                    document.getElementById("p3_2").innerHTML="      In the field of biomanufacturing, the selection of different chassis cells plays a key role in the biosynthesis of terpenoids. IPP and DMAPP, the precursors of terpenoids in E. coli, are mainly used for prenylation of tRNA and synthesis of FPP, and then for synthesis of quinones and cell walls[3]. Therefore, these chassis cells are mainly used for biosynthesis of alcohols and acids rather than terpenoids. In addition, due to the lack of post-translational modification, it is difficult for bacteria to express cytochrome P450, and many terpenoids have antibacterial activity. However, <i>Saccharomyces cerevisiae</i> has been widely used in terpenoid biosynthesis because it is generally regarded as safe (GRAS),10 industrially robust and able to functionally express eukaryotic cytochrome P450 enzymes[4]. Therefore, SCUT-China will also consider using <i>S. cerevisiae</i> to build our Nootkatone cell factory.";
 +
                    document.getElementById("p3_4").innerHTML="    After constructing a <i>saccharomyces cerevisiae</i> cell factory capable of producing Nootkatone, we found that the construction of metabolic pathways usually involves the expression of multiple genes, whose expression levels span several orders of magnitude, and the regulation of gene expression by endogenous gene regulatory elements alone is not sufficient[5]. In addition, we also found that CnVS is the limiting enzyme in the metabolic pathway of Nootkatone synthesis, and its expression level plays a key role in Nootkatone production[2]. How to effectively improve the expression level of VS to improve Nootkatone precursor (valencene) and Nootkatone production is a big problem we are facing.<br><br>To solve the above two problems, we turned our attention to promoters that control gene transcription[6]. As the initiation of gene transcription, promoters are key components of cell factory design and metabolic pathway modification. Due to the limited number of well-characterized promoters in yeast and their small dynamic ranges, it is often difficult to satisfy the fine regulation of genes, optimize metabolic flux, and improve the yield of the target product[7]. Therefore, SCUT-China wants to obtain high-strength promoters to increase the expression of CnVS through engineering modification of <i>S. cerevisiae</i> natural promoters, and to obtain a good dynamic promoter library, which will provide useful and efficient help for the improvement of metabolic flux optimization target product yield and the design of cell factory.<br><br>In labs, our promoter engineering will be carried out from <a href='https://2021.igem.org/Team:SCUT-China/Design'>Wet Lab </a>and <a href='https://2021.igem.org/Team:SCUT-China/Model'>Dry Lab</a> respectively. Wet Lab mainly focuses on UAS combining different promoters to construct a series of hybrid-promoters with different strengths. Dry Lab obtained promoters with different intensities mainly from nucleosome affinity of promoters.";
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Revision as of 00:19, 22 October 2021


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Overview

A group of students from SCUT in Guangzhou came up with an idea to alleviate the scourge of mosquitoes. In inspiration, we skillfully used Baidu index big data analysis, investigated the official data of the WORLD Health Organization and EPA, and known a new mosquito repellent ingredient--Nootkatone. In the conception stage of our project, based on the rise of biological manufacturing and synthetic biology, we considered from three aspects: the selection of chassis cells, the identification of synthetic pathways and how to increase yield, so as to build a cell factory with high yield of Nootkatone and to realize our Nootka-Boom!

此处为pdf窗口示例

A group of students from SCUT in Guangzhou came up with an idea to alleviate the scourge of mosquitoes. In inspiration, we skillfully used Baidu index big data analysis, investigated the official data of the WORLD Health Organization and EPA, and known a new mosquito repellent ingredient--Nootkatone. In the conception stage of our project, based on the rise of biological manufacturing and synthetic biology, we considered from three aspects: the selection of chassis cells, the identification of synthetic pathways and how to increase yield, so as to build a cell factory with high yield of Nootkatone and to realize our Nootka-Boom!

Inspiration

Regional necessity:

South China University of Technology is located in the humid and rainy Guangzhou, where the warm climate with unclear seasons is not only suitable for human to live but also for mosquito breeding. Whether indoors or outdoors, the mosquito problem has always plagued us, as well as other people living in Guangzhou. According to the data of Baidu index, from 2013 to 2021, the users who search the keyword of mosquito repellent on the Internet are most from Guangdong. And in Guangdong, the most dominant users having searched relevant questions are from Guangzhou. This can show that the mosquito problem is a high concern and urgent problem for people around us.

    < >

    Image Source:baidu.com

    International necessity:

    Mosquito bites bring not only itchy skin, but also the risk of contracting infections mosquito-borne diseases. Mosquitoes are vector organisms and diseases that use mosquitoes as vectors include yellow fever, dengue fever, epidemic B encephalitis, malaria, lymphatic filariasis, Chikungunya, West Nile virus disease, and Rift Valley fever. In the case of dengue fever, for example, WHO listed it as one of the top 10 threats to global health in 2019. About 390 million people in 128 countries are at risk of dengue fever, with an average of 96 million people infected each year, including asymptomatic carriers. The mortality rate has remained stable at around 0.12%, which means that thousands of people die each year from dengue infection.

    Data Source: World Health Organization

    The most direct solution to the problem seems to be the use of insecticide. However, monitoring has found that insecticide resistance is widespread among malaria vectors. According to the World Health Organization, 65 of 89 malaria-endemic countries have reported at least one endemic vector resistant to pyrethroids since 2010. All four classes of insecticides currently used for adult malaria vector control, pyrethroids, organochlorines, carbamates and organophosphates, have spread widely in Africa, the Americas, Southeast Asia, the eastern Mediterranean and the Western Pacific. Moreover, malaria vectors in some parts of Africa are now able to survive in environments with high concentrations of insecticides, suggesting that resistance in some malaria vector organisms is now increasing over time.

    Source: WHO Malaria Threats Map.

    In the EPA's report on new repellent ingredients, we noted that compared with existing repellent ingredients, Nootkatone can maintain high repellent effect stably for a long time. As proven by EPA's data, the high-purity Nootkatone not only has high safety, but also has no irritation, no skin sensitization, and is a qualified mosquito repellent ingredient. According to the EPA, the creation of a new effective insecticide ingredient,Nootkatone, will help address the growing insecticide resistance to other products currently in use. However, currently there is no popular product related to Nootkatone on the market because its current primary production method is plant-based extraction, which is expensive and difficult to produce in large quantities. In order to further enable the wider application of Nootkatone, to exploit its value and to explore more of its possibilities, we have undertaken our project - Nootka-Boom!

    Nootka-Boom！

    Source of project

    Synthetic biology with engineering design concept, design and transformation of organism to have a goal, subvert the biological technique innovation, is expected to decode the human resources, environment and other fields important challenge to provide a new solution With the rapid development of synthetic biology, through the microbial production of high value-added compounds become attractive In April 2013,Nature published a study on artemisinin, a highly effective semi-synthetic antimalarial drug in Saccharomyces cerevisiae, which greatly promoted the industrialization of microbial semi-synthetic artemisinin. This is the most successful case of heterologous synthesis of high value-added products by microorganisms.

    Nootkatone is a natural ingredient found in some Alaskan yellow cedar herbs and citrus fruits that is responsible for grapefruit's distinctive smell. Nootkatone has long been used in many food products that have a citrus flavor or smell. DVBD scientists have found that Nootkatone is an effective mosquito repellent and insecticide that can be used against mosquito ticks and other pests[1].

    There are three main production methods of Nootkatone: Plant extraction method, chemical synthesis and microbial synthesis. Terpenoids content in plants is usually very low, the extraction method is easy to cause serious damage to wild plant resources, and the planting and processing of its crops are affected by many factors such as climate, environment and transportation, so the yield, quality and price of Nootkatone have uncontrollable changes, and the economic feasibility is poor. At present, the industry is often through a precursor of the direct oxidation of price is relatively cheap Valencene to obtain Nootkatone. But the reaction usually involves some environmental unfriendly oxidant, such as heavy metal salts. By contrast, microbial synthesis method is not restricted by raw material Green clean production process, has a great advantage[2]. Such as the Nature The focus of the synthetic biology industry is rapidly shifting to the over $20 billion fine chemicals market, which will rapidly advance the heterologous biosynthesis of natural products Therefore, to meet the growing demand for Nootkatone, SCUT-China wants to use synthetic biology methods to create a cell factory with high-yield of Nootkatone.

    Selection of chassis cell

    In the field of biomanufacturing, the selection of different chassis cells plays a key role in the biosynthesis of terpenoids. IPP and DMAPP, the precursors of terpenoids in E. coli, are mainly used for prenylation of tRNA and synthesis of FPP, and then for synthesis of quinones and cell walls[3]. Therefore, these chassis cells are mainly used for biosynthesis of alcohols and acids rather than terpenoids. In addition, due to the lack of post-translational modification, it is difficult for bacteria to express cytochrome P450, and many terpenoids have antibacterial activity. However, Saccharomyces cerevisiae has been widely used in terpenoid biosynthesis because it is generally regarded as safe (GRAS),10 industrially robust and able to functionally express eukaryotic cytochrome P450 enzymes[4]. Therefore, SCUT-China will also consider using S. cerevisiae to build our Nootkatone cell factory.

    Identification of biosynthetic pathways

    All terpenes are biosynthesized through mevalonate pathway (MVA) in yeast, originating from acetyl-CoA. And the intermediate product farnesyl diphosphate (FPP) is the direct precursor of valencene, which is catalyzed by valencene synthase (VS) to synthesize valencene. Then, valencene is oxidated to β-Nootkatol and (+)-nootkatone by HPO, AtCPR and ADH[2]. SCUT-China will introduce the Nootkatone synthesis pathway into S. cerevisiae to build our cell factory.

    Idea to realize Nootka-Boom!

    After constructing a saccharomyces cerevisiae cell factory capable of producing Nootkatone, we found that the construction of metabolic pathways usually involves the expression of multiple genes, whose expression levels span several orders of magnitude, and the regulation of gene expression by endogenous gene regulatory elements alone is not sufficient[5]. In addition, we also found that CnVS is the limiting enzyme in the metabolic pathway of Nootkatone synthesis, and its expression level plays a key role in Nootkatone production[2]. How to effectively improve the expression level of VS to improve Nootkatone precursor (valencene) and Nootkatone production is a big problem we are facing.

    To solve the above two problems, we turned our attention to promoters that control gene transcription[6]. As the initiation of gene transcription, promoters are key components of cell factory design and metabolic pathway modification. Due to the limited number of well-characterized promoters in yeast and their small dynamic ranges, it is often difficult to satisfy the fine regulation of genes, optimize metabolic flux, and improve the yield of the target product[7]. Therefore, SCUT-China wants to obtain high-strength promoters to increase the expression of CnVS through engineering modification of S. cerevisiae natural promoters, and to obtain a good dynamic promoter library, which will provide useful and efficient help for the improvement of metabolic flux optimization target product yield and the design of cell factory.

    In labs, our promoter engineering will be carried out from Wet Lab and Dry Lab respectively. Wet Lab mainly focuses on UAS combining different promoters to construct a series of hybrid-promoters with different strengths. Dry Lab obtained promoters with different intensities mainly from nucleosome affinity of promoters.

    References

    1. Clarkson, T.C., et al., Nootkatone Is an Effective Repellent against Aedes aegypti and Aedes albopictus. INSECTS, 2021. 12(5).

    2. Ouyang, X., et al., Stepwise engineering of Saccharomyces cerevisiae to produce (+)-valencene and its related sesquiterpenes. RSC ADVANCES, 2019. 9(52): p. 30171-30181.

    3. Wang, D., Z. Dai, and X. Zhang, [Production of plant-derived natural products in yeast cells - A review]. Wei sheng wu xue bao = Acta microbiologica Sinica, 2016. 56(3): p. 516-29.

    4. Dai, Z.B., et al., Yeast synthetic biology for high-value metabolites. Fems Yeast Research, 2015. 15(1): p. 11.

    5. Chen, X., et al., DCEO Biotechnology: Tools To Design, Construct, Evaluate, and Optimize the Metabolic Pathway for Biosynthesis of Chemicals. Chemical Reviews, 2018. 118(1): p. 4-72.

    6. Young, E. and H. Alper, Synthetic Biology: Tools to Design, Build, and Optimize Cellular Processes. Journal of Biomedicine and Biotechnology, 2010.

    7. Maury, J., et al., Reconstruction of a bacterial isoprenoid biosynthetic pathway in Saccharomyces cerevisiae. Febs Letters, 2008. 582(29): p. 4032-4038