(Blanked the page) |
|||
Line 1: | Line 1: | ||
+ | {{NPU-China/mmp}} | ||
+ | <html lang="en"> | ||
+ | <head> | ||
+ | <!-- Bootstrap Core CSS --> | ||
+ | <link href="https://cdn.bootcss.com/bootstrap/3.3.7/css/bootstrap.min.css" rel="stylesheet"> | ||
+ | |||
+ | <!-- Custom CSS --> | ||
+ | <link href="https://2017.igem.org/Template:NPU-China/css?action=raw&ctype=text/css" rel="stylesheet"> | ||
+ | |||
+ | <!-- Custom Fonts --> | ||
+ | <link href="https://cdn.bootcss.com/font-awesome/4.7.0/css/font-awesome.min.css" rel="stylesheet"> | ||
+ | |||
+ | <!-- jQuery --> | ||
+ | <script src="https://cdn.bootcss.com/jquery/3.2.1/jquery.min.js"></script> | ||
+ | |||
+ | <!-- Bootstrap Core JavaScript --> | ||
+ | <script src="https://cdn.bootcss.com/bootstrap/3.3.7/js/bootstrap.min.js"></script> | ||
+ | |||
+ | <!-- HTML5 Shim and Respond.js IE8 support of HTML5 elements and media queries --> | ||
+ | <!-- WARNING: Respond.js doesn't work if you view the page via file:// --> | ||
+ | <!--[if lt IE 9]> | ||
+ | <script src="https://oss.maxcdn.com/libs/html5shiv/3.7.0/html5shiv.js"></script> | ||
+ | <script src="https://oss.maxcdn.com/libs/respond.js/1.4.2/respond.min.js"></script> | ||
+ | <![endif]--> | ||
+ | <style> | ||
+ | </style> | ||
+ | </head> | ||
+ | |||
+ | |||
+ | <body> | ||
+ | |||
+ | <!-- Navigation --> | ||
+ | <nav class="navbar navbar-inverse navbar-fixed-top" role="navigation"> | ||
+ | <div class="container"> | ||
+ | <div class="navbar-header"> | ||
+ | <button type="button" class="navbar-toggle" data-toggle="collapse" data-target="#bs-example-navbar-collapse-1"> | ||
+ | <span class="sr-only">Toggle navigation</span> | ||
+ | <span class="icon-bar"></span> | ||
+ | <span class="icon-bar"></span> | ||
+ | <span class="icon-bar"></span> | ||
+ | </button> | ||
+ | <a class="navbar-brand" href="https://2017.igem.org/Team:NPU-China"> | ||
+ | <img src="https://static.igem.org/mediawiki/2017/2/29/NPU-logo.png" style="max-width:50px;margin-top:-10px;"> | ||
+ | </a> | ||
+ | </div> | ||
+ | <div class="collapse navbar-collapse" id="bs-example-navbar-collapse-1"> | ||
+ | <ul class="nav navbar-nav navbar-right"> | ||
+ | <li> | ||
+ | <a href="https://2017.igem.org/Team:NPU-China">Home</a> | ||
+ | </li> | ||
+ | <li class="dropdown"> | ||
+ | <a href="#" class="dropdown-toggle" data-toggle="dropdown">Team | ||
+ | <b class="caret"></b> | ||
+ | </a> | ||
+ | <ul class="dropdown-menu"> | ||
+ | <li> | ||
+ | <a href="https://2017.igem.org/Team:NPU-China/Aboutus">About us</a> | ||
+ | </li> | ||
+ | <li> | ||
+ | <a href="https://2017.igem.org/Team:NPU-China/Attributions">Attributions</a> | ||
+ | </li> | ||
+ | </ul> | ||
+ | </li> | ||
+ | <li class="dropdown active"> | ||
+ | <a href="#" class="dropdown-toggle" data-toggle="dropdown">Project | ||
+ | <b class="caret"></b> | ||
+ | </a> | ||
+ | <ul class="dropdown-menu"> | ||
+ | <li> | ||
+ | <a href="https://2017.igem.org/Team:NPU-China/Background">Background</a> | ||
+ | </li> | ||
+ | <li> | ||
+ | <a href="https://2017.igem.org/Team:NPU-China/Description">Description</a> | ||
+ | </li> | ||
+ | <li> | ||
+ | <a href="https://2017.igem.org/Team:NPU-China/Design">Design</a> | ||
+ | </li> | ||
+ | <li> | ||
+ | <a href="https://2017.igem.org/Team:NPU-China/Model">Model</a> | ||
+ | </li> | ||
+ | <li> | ||
+ | <a href="https://2017.igem.org/Team:NPU-China/Proofofconcept">Proof of concept</a> | ||
+ | </li> | ||
+ | <li> | ||
+ | <a href="https://2017.igem.org/Team:NPU-China/Demonstrate">Demonstrate</a> | ||
+ | </li> | ||
+ | </ul> | ||
+ | </li> | ||
+ | <li class="dropdown"> | ||
+ | <a href="#" class="dropdown-toggle" data-toggle="dropdown">Parts | ||
+ | <b class="caret"></b> | ||
+ | </a> | ||
+ | <ul class="dropdown-menu"> | ||
+ | <li> | ||
+ | <a href="https://2017.igem.org/Team:NPU-China/BasicParts">Basic Parts</a> | ||
+ | </li> | ||
+ | <li> | ||
+ | <a href="https://2017.igem.org/Team:NPU-China/CompositeParts">Composite Parts</a> | ||
+ | </li> | ||
+ | </ul> | ||
+ | </li> | ||
+ | <li> | ||
+ | <a href="https://2017.igem.org/Team:NPU-China/Hardware">Hardware</a> | ||
+ | </li> | ||
+ | <li class="dropdown"> | ||
+ | <a href="#" class="dropdown-toggle" data-toggle="dropdown">HP | ||
+ | <b class="caret"></b> | ||
+ | </a> | ||
+ | <ul class="dropdown-menu"> | ||
+ | <li> | ||
+ | <a href="https://2017.igem.org/Team:NPU-China/HP/Silver">Silver</a> | ||
+ | </li> | ||
+ | <li> | ||
+ | <a href="https://2017.igem.org/Team:NPU-China/HP/Gold_Integrated">Gold</a> | ||
+ | </li> | ||
+ | </ul> | ||
+ | </li> | ||
+ | <li> | ||
+ | <a href="https://2017.igem.org/Team:NPU-China/Collaborations">Collaborations</a> | ||
+ | </li> | ||
+ | <li> | ||
+ | <a href="https://2017.igem.org/Team:NPU-China/Achievements">Achievements</a> | ||
+ | </li> | ||
+ | <li> | ||
+ | <a href="https://2017.igem.org/Team:NPU-China/InterLab">InterLab</a> | ||
+ | </li> | ||
+ | |||
+ | <li class="dropdown"> | ||
+ | <a href="#" class="dropdown-toggle" data-toggle="dropdown">Notebook | ||
+ | <b class="caret"></b> | ||
+ | </a> | ||
+ | <ul class="dropdown-menu"> | ||
+ | <li> | ||
+ | <a href="https://2017.igem.org/Team:NPU-China/Labnotes">Labnotes</a> | ||
+ | </li> | ||
+ | <li> | ||
+ | <a href="https://2017.igem.org/Team:NPU-China/Protocols">Protocols</a> | ||
+ | </li> | ||
+ | </ul> | ||
+ | </li> | ||
+ | </ul> | ||
+ | </div> | ||
+ | </div> | ||
+ | </nav> | ||
+ | |||
+ | <!-- Page Content --> | ||
+ | <div class="batu" style="background: url('https://static.igem.org/mediawiki/2017/f/fe/Npu-background.png') no-repeat fixed; overflow: hidden;"> | ||
+ | <img class="img-responsive" src="https://static.igem.org/mediawiki/2017/9/97/%E9%A2%98%E7%9B%AE%E9%80%9A%E6%A0%8Fdescription.jpg"> | ||
+ | <div class="container" style="padding-top:70px"> | ||
+ | <div class="row"> | ||
+ | <div class="col-md-12"> | ||
+ | <h4>This year, we focus on an important chemical organic synthesis raw material – acrylic acid. We hope to | ||
+ | build an efficient cell factory to achieve "all green" production of acrylic acid efficiently, so | ||
+ | we choose glycerol as the raw material for microbial cell factories to produce bulk chemical products. | ||
+ | Glycerol has the advantage of being cheap and environmental-friendly, and allaying the pressure on | ||
+ | the by-product waste in the production of biodiesel. In addition, compared to glucose, xylose and | ||
+ | other carbohydrate substrates, glycerol metabolism can produce higher reducing power. | ||
+ | <br><br> Complex synthetic pathway, vague synthetic mechanism and low efficiency of the synthesis, are the | ||
+ | current shortcomings of acrylic biosynthesis method. How to build a short and efficient acrylic biosynthetic | ||
+ | pathway to build a highly efficient acrylic biosynthetic factory is the key to success! This is also | ||
+ | the entry point for our project this year. | ||
+ | <br><br> In the previous experiments, we further demonstrated the new function of ceaS2 enzyme, which can | ||
+ | catalyze the production of acrylic acid with DHAP (hydroxyxy acetone phosphate) or G3P (glyceraldehyde | ||
+ | 3-phosphate) as substrate. Because acrylic acid is not the main product of ceaS2 enzyme, the catalytic | ||
+ | effect of wild-type ceaS2 enzyme is very weak and the yield of acrylic acid is only 1mg / L. So we | ||
+ | carried out engineering modification of this enzyme to improve the catalytic effect of the core part. | ||
+ | We used the AEMD (Auto Enzyme Mutation Design) platform to identify mutational sites and screened | ||
+ | for high catalytic efficiency by HPLC (High Performance Liquid Chromatography) (Learn more about | ||
+ | HPLC!) and HTS (High throughput screening) of the ceaS2 mutant. | ||
+ | <br><br> In the selection of the chassis organisms, we chose E. coli which is a kind of classic chassis organism | ||
+ | in prokaryote and Saccharomyces cerevisiae which is the most easily manipulated chassis organism | ||
+ | in eukaryotes. | ||
+ | <br><br> The GDC (GlyDH-DAK-Ceas2) pathway was designed in order to improve the ability of chassis cells | ||
+ | to convert glycerol to DHAP and finally synthesize acrylic acid. We also added NOX (NADH dehydrogenase) | ||
+ | and CAT (Catalase) to this pathway to provide the required reducing power for GLYDH by two layers | ||
+ | of substrate circulation. Ultimately, GNCDC (GlyDH-NOX-CAT-DAK-ceaS2) is our desired new biosynthetic | ||
+ | pathway of acrylic acid. | ||
+ | <br><br> In order to determine the effect of the new route on the yield of acrylic acid before the wet experiment | ||
+ | and to analyze the stability and robustness of the new route, we asked SCAU-China to help us did | ||
+ | the work of metabolic flow modeling. We analyzed the changes of carbon flux of the two key intermediates, | ||
+ | DHAP and G3P, before and after joining the new pathway. The modeling results show that the new routes | ||
+ | can increase the carbon flow of DHAP and G3P, thereby contributing to the increase in the production | ||
+ | of acrylic acid. | ||
+ | <br><br> We used whole cell catalysis to carry out the initial acrylic acid synthesis "fermentation" process. | ||
+ | The advantage of whole cell catalysis is that the intracellular complete multi-enzyme system can | ||
+ | achieve the cascade reaction of enzyme, so as to make up the deficiency of cascade reaction in reaction | ||
+ | which only use pure enzyme and improve the catalytic efficiency. While eliminating the complex process | ||
+ | in enzyme purification, it is easier to carry out the reaction and lower production costs. | ||
+ | <br><br> We optimized the reaction process, selected the carbon source, buffer, temperature, pH, reaction | ||
+ | time and other conditions to optimize the production process of the cell factory. | ||
+ | <br><br> We also made Hardware (Click Here) to simulate the industrial production process of acrylic acid!</h4> | ||
+ | |||
+ | </div> | ||
+ | |||
+ | </div> | ||
+ | <!-- Blog Post Row --> | ||
+ | |||
+ | |||
+ | </div> | ||
+ | <img src="https://static.igem.org/mediawiki/2017/0/0c/Jz.png" class="img-responsive"> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | </body> | ||
+ | |||
+ | </html> |
Revision as of 19:22, 31 October 2017