Difference between revisions of "Team:ShanghaiFLS China"

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                     <p>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. This year, we modified the metabolic pathway of <em>Pichia Pastoris</em>, a type of yeast that can convert methanol to other carbon compounds by re-engineering its homogeous circuits expressing the transcription factors that would up-regulate the expression of AOX1, the protein allowing it to metabolize methanol. Our experiments yielded positive results. In addition, we integrated human practices into our project by talking to professionals and distributing questionnaires to more than 700 individuals living in Shanghai. We designed two models to make sure our experiment design was valid and to prove its industrial uses. Want to know more? Let's start the journey!</p>
 
                     <p>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. This year, we modified the metabolic pathway of <em>Pichia Pastoris</em>, a type of yeast that can convert methanol to other carbon compounds by re-engineering its homogeous circuits expressing the transcription factors that would up-regulate the expression of AOX1, the protein allowing it to metabolize methanol. Our experiments yielded positive results. In addition, we integrated human practices into our project by talking to professionals and distributing questionnaires to more than 700 individuals living in Shanghai. We designed two models to make sure our experiment design was valid and to prove its industrial uses. Want to know more? Let's start the journey!</p>
 
                     <a href="https://2019.igem.org/Team:ShanghaiFLS_China/Team"> Team Members </a>  
 
                     <a href="https://2019.igem.org/Team:ShanghaiFLS_China/Team"> Team Members </a>  

Revision as of 14:21, 19 October 2019

ShanghaiFLS_China: Home

Experiments

The Optimization of the Metabolic Pathways of P. pastoris in Medicine Production via Methanol Fermentation

Methanol is a major byproduct of the coal industry. Engineered Pichia pastoris GS115, a strain of methylotrophic yeast, is capable of converting methanol into medical compounds such as the insulin precursor and lovastatin. However, in such P. pastoris, the metabolism of methanol is highly specific and results in significant oxygen consumption and heat generation, which have limited its industrial applications. We aim to address this issue by improving the methanol conversion rate in P. pastoris by re-engineering its homogeneous circuits expressing the transcription factors that would up-regulate the expression of AOX1, the protein allowing it to metabolize methanol. Through our multiple rounds of experiments, we eventually acquired strains that are capable of yielding an up to 20% increase in total GFP production per gram methanol compared to the wildtype. Based on our modeling, such strains should have an (percent) decrease in heat generation per unit product and a (percent) decrease in oxygen consumption compared to the wildtype.

Design Experiment Results Parts

Outside the Lab

Demonstrate

(hyfff)

Model

To get an idea of how much more efficient our constructs will be compared with the wild-type P. pastoris, 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.

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 will be difficult and decided to use a model to demonstrate its effectiveness.