Difference between revisions of "Team:ShanghaiFLS China"

Revision as of 06:59, 21 October 2019

ShanghaiFLS_China: Home

Pichia Pastoris GS115:

The Methanol Converter

From methanol to medicine, how to do it better? GS115:

Methanol is a major byproduct of the coal industry. Engineered Pichia pastoris GS115 can convert methanol into medical compounds such as the insulin precursor and lovastatin. However, this pathway results in significant oxygen consumption and heat generation, limiting its industrial applications. We improved the methanol conversion rate in P. pastoris by rewiring its homogeneous circuits expressing the transcription factors that would up-regulate the expression of alcohol oxidase 1, the protein that allows it to metabolize methanol. Through multiple rounds of experiments, we acquired strains that are able to yield an up to 20% increase in total production per unit methanol compared to the wildtype. In other words, these strains generate less heat producing the same amount of produce.

Design Experiment Results Parts

About us

We are a diverse group of high school students from China, studying in Beijing, Tianjin, the United States or Switzerland, as well as in Shanghai Foreign Language School. We have different hobbies, ambitions, and views towards bioengineering. We also have different roles to play this fascinating project. For more information about us, please check the links below:

Team Members Attributions

Outside the Lab

Demonstrate

When we asked research professionals and industrial leaders whether our project would be successful in the industry, they were not optimistic. To address this doubt, we constructed a model calculating the oxygen consumption and heat generation of our yeast strains, two main factors to be considered in industrial processes. We based our models on validated and published equasions, and when we looked at the results of our calculations, they all prove adequate possibilities of industrial production (yes!).

Model

While the model above is constructed after our experiments, before we even got into the lab, we wanted to get an idea of how much more efficient our constructs will be compared with the wild-type P. pastoris. So we modeled our constructs based on various statistics published in Wang et al., 2016, Shi et al., 2019 and Liang et al., 2012, and calculated basic parameters for each promoter and transcription factor in our three constructs. Specifically, we modulated the activity of one promoter under the effect of one unit transcription factor, and used it to calculate the overall expression efficiency of our constructs. As it turned out, the results looked pretty encouraging, and so we proceeded with our experiment.

Human Practices

From the start of our project, we actively consulted peers, research professionals, industry experts, and the general public for advice and guidance. We first met up with Dr. Cai from ECUST, who introduced us to Pichia pastoris GS115, which we decided to base our project upon. We also interviewed two industry leaders. One of them, Dr. Hu, CEO of GTL Biotech, illustrated to us the great potential of methanol as a carbon source, which happens to be what P. pastoris is tailored to metabolize. After designing our experiments, we referred to the opinion of the public for evaluations of acceptability, feasibility and general impact via a questionnaire. We also became aware that industrializing our project design might be difficult, so decided to use a model to evaluate its effectiveness.

@@ Line 105: / Line 105: @@
                      <h3 class="editContent"><em>Pichia Pastoris</em> GS115:</h3>
                      <h2 class="editContent">The Methanol Converter</h2>
-                     <p>Methanol is a major byproduct of the coal industry. Engineered <em>Pichia pastoris</em> GS115 can convert methanol into medical compounds such as the insulin precursor and lovastatin. However, this pathway is highly specific and results in significant oxygen consumption and heat generation, limiting its industrial applications. We improved the methanol conversion rate in <em>P. pastoris</em> by rewiring its homogeneous circuits expressing the transcription factors that would up-regulate the expression of AOX1, the protein that allows it to metabolize methanol. Through multiple rounds of experiments, we acquired strains that are able to yield an up to 20% increase in total production per gram methanol compared to the wildtype.</p>
+                    <h3 class="editContent"><em>From methanol to medicine, how to do it better?</em> GS115:</h3>
+                     <p>Methanol is a major byproduct of the coal industry. Engineered <em>Pichia pastoris</em> GS115 can convert methanol into medical compounds such as the insulin precursor and lovastatin. However, this pathway results in significant oxygen consumption and heat generation, limiting its industrial applications. We improved the methanol conversion rate in <em>P. pastoris</em> by rewiring its homogeneous circuits expressing the transcription factors that would up-regulate the expression of alcohol oxidase 1, the protein that allows it to metabolize methanol. Through multiple rounds of experiments, we acquired strains that are able to yield an up to 20% increase in total production per unit methanol compared to the wildtype. In other words, these strains generate less heat producing the same amount of produce.</p>
                  <a href="https://2019.igem.org/Team:ShanghaiFLS_China/Design">Design</a>
                  <a href="https://2019.igem.org/Team:ShanghaiFLS_China/Experiments">Experiment</a>
@@ Line 122: / Line 123: @@
          <h3 class="heading">About us</h3>
          <div class="row about-grids">
-                 <p class="my-4">We are a diverse group of high school students from China, studying in Beijing, Tianjin, the United States or Switzerland, as well as in Shanghai Foreign Language School. We have different hobbies, ambitions, and views towards bioengineering. We also have different jobs o realize this complicated project. For more information about us, please click on the links below:<br /><br /></a>
+                 <p class="my-4">We are a diverse group of high school students from China, studying in Beijing, Tianjin, the United States or Switzerland, as well as in Shanghai Foreign Language School. We have different hobbies, ambitions, and views towards bioengineering. We also have different roles to play this fascinating project. For more information about us, please check the links below:<br /><br /></a>
                  <a href="https://2019.igem.org/Team:ShanghaiFLS_China/Team"> Team Members </a>
                  <a href="https://2019.igem.org/Team:ShanghaiFLS_China/Attributions"> Attributions </a>
@@ Line 138: / Line 139: @@
              <div class="col-lg-4 col-md-6 w3layouts w3-icon-grid1">
                  <h3><span class="fa mr-2 fa-cogs" aria-hidden="true"></span><a href="https://2019.igem.org/Team:ShanghaiFLS_China/demonstrate"><u>Demonstrate</u></a></h3>
-                 <p>When we asked professors and industrial leaders whether our project would be successful in the industry, they were not optimistic. However, due to constraints in our experimental settings, we were not able to demonstrate through experiments how our designs would work. This problem led us to construct a model, modifying existent formulas from literary works, to demonstrate this possibility. The rate of oxygen consumption and heat emission are two main factors to be considered in industrial processes, and our project mainly looks at methanol consumption. When we looked at the three factors separately in three models below, they all prove adequate possibilities of industrial production.</p>
+                 <p>When we asked research professionals and industrial leaders whether our project would be successful in the industry, they were not optimistic. To address this doubt, we constructed a model calculating the oxygen consumption and heat generation of our yeast strains, two main factors to be considered in industrial processes. We based our models on validated and published equasions, and when we looked at the results of our calculations, they all prove adequate possibilities of industrial production (yes!).</p>
              </div>
              <div class="col-lg-4 col-md-6 w3layouts w3-icon-grid1">
                  <h3><span class="fa mr-2 fa-laptop" aria-hidden="true"></span><a href="https://2019.igem.org/Team:ShanghaiFLS_China/model"><u>Model</u></a></h3>
-                 <p>To get an idea of how much more efficient our constructs will be compared with the wild-type <em>P. pastoris</em>, we modeled our constructs based on various statistics published in Wang et al., 2016, Shi et al., 2019 and Liang et al., 2012. Based on these data, we calculated basic parameters for each promoter and transcription factor in our three constructs, specifically, the activity of a promoter under the effect of 1 unit transcription factor. As it turned out, we relatively successfully calculated the eventual results of our experiment.</p>
+                 <p>While the model above is constructed after our experiments, before we even got into the lab, we wanted to get an idea of how much more efficient our constructs will be compared with the wild-type <em>P. pastoris</em>. So we modeled our constructs based on various statistics published in Wang et al., 2016, Shi et al., 2019 and Liang et al., 2012, and calculated basic parameters for each promoter and transcription factor in our three constructs. Specifically, we modulated the activity of one promoter under the effect of one unit transcription factor, and used it to calculate the overall expression efficiency of our constructs. As it turned out, the results looked pretty encouraging, and so we proceeded with our experiment.</p>
              </div>
              <div class="col-lg-4 col-md-6 w3layouts w3-icon-grid1">
                  <h3><span class="fa mr-2 fa-camera" aria-hidden="true"></span><a href="https://2019.igem.org/Team:ShanghaiFLS_China/Human_Practices"><u>Human Practices</u></a></h3>
-                 <p>From the start of our project, we actively consulted peers, research professionals, industry experts, and the general public for advice and guidance. We first met up with Dr. Cai from ECUST, who introduced us to <em>Pichia pastoris GS115</em>, which we decided to base our project upon. We also interviewed two industry leaders. One of them, Dr. Hu, CEO of <em>GTL Biotech</em>, illustrated to us the great potential of methanol as a carbon source, which happens to be what <em>P. pastoris</em> is tailored to metabolize. After designing our experiments, we referred to the opinion of the public for evaluations of acceptability, feasibility and general impact via a questionnaire. We also became aware that industrializing our project design will be difficult and decided to use a model to demonstrate its effectiveness.</p>
+                 <p>From the start of our project, we actively consulted peers, research professionals, industry experts, and the general public for advice and guidance. We first met up with Dr. Cai from ECUST, who introduced us to <em>Pichia pastoris GS115</em>, which we decided to base our project upon. We also interviewed two industry leaders. One of them, Dr. Hu, CEO of <em>GTL Biotech</em>, illustrated to us the great potential of methanol as a carbon source, which happens to be what <em>P. pastoris</em> is tailored to metabolize. After designing our experiments, we referred to the opinion of the public for evaluations of acceptability, feasibility and general impact via a questionnaire. We also became aware that industrializing our project design might be difficult, so decided to use a model to evaluate its effectiveness.</p>
          </div>
      </div>