Difference between revisions of "Team:ASIJ Tokyo/Description"

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Our iGEM project was inspired by the results of a collaborative study between Keio University and Kyoto Institute of Technology, two prestigious Japanese universities. Their research investigated the ability of PETase, an enzyme, to degrade PET plastic using a bacterium known as <i>Ideonella Saikainesis</i>. The success of PETase in the degradation of PET plastic into Polyethylene Terephthalate and Ethylene Glycol sparked our interest in improving the Japanese ecological environment. Thus, the goal of our iGEM project is to create a biobrick incorporating an ideal Andersen promoter to optimize the use of PETase in an <i>E. coli</i> bacteria system. Past iGEM projects centered on plastic degradation, such as those of Turkey 2014, University of Washington 2012 and Darmstadt 2012, also provided us with inspiration on the potential applications for our project, as well as lab protocols. </h4>
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Our iGEM project was inspired by the results of a collaborative study between Keio University and Kyoto Institute of Technology, two prestigious Japanese universities. Their research investigated the ability of PETase, an enzyme, to degrade PET plastic using a bacterium known as <i>Ideonella Saikainesis</i>. The success of PETase in the degradation of PET plastic into Polyethylene Terephthalate and Ethylene Glycol sparked our interest in improving the Japanese ecological environment. Thus, the goal of our iGEM project is to create a biobrick incorporating an ideal Anderson promoter to optimize the use of PETase in an <i>E. coli</i> bacteria system. Past iGEM projects centered on plastic degradation, such as those of Turkey 2014, University of Washington 2012 and Darmstadt 2012, also provided us with inspiration on the potential applications for our project, as well as lab protocols. </h4>
  
 
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The goal of our iGEM project is to create an optimal PETase biobrick that will be available to future iGEM teams for use in plastic degradation. We hoped that this will make experiments in the following years easier to conduct. However, in order to even get to that stage, we recognized that we needed a well thought out procedure. Our team is currently working on this step. Thus far we have determined that we will be using a western blot test to help us deal with the signal peptides. We also have three promoters (one on the slightly weaker side and two on the stronger side) taken from the list of Anderson promoters to test in order to determine how to make PETase most efficient. <br><br>
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Our team focused on making a biobrick featuring the PETase enzyme to contribute to the IGEM registry. In addition, we hoped to optimize the initial step of PET breakdown, or depolymerization, so as to expedite the overall process of degradation. To accomplish these goals, we selected several Anderson promoters of different strengths (weak, moderately-strong, and strong) to test for optimizing PETase production. The faster the rate of transcription for the PETase gene, the more PETase enzyme is produced. The greater the volume of enzyme, the faster the rate of depolymerization, since there is more enzyme available to ‘work’ on a plastic sample.<br><br>
  
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However, before testing such promoters, we need to isolate and evaluate each promoters' effectiveness and efficiency. This information is essential before we proceed onto our next steps and think about real world applications. Questions we may ask ourselves following this first step include: “How long will the degradation take?” and “Will the byproducts of the degradation be harmful in any way to the environment or to people?” <br><br>
 
  
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Our aim, as stated above, is to find a faster, more efficient process to degrade plastic. Answers to such questions will be essential in seeking solutions to global environmental problems. In regards to the first question, to our current knowledge, 450 years is the timeframe for the natural biodegradation of PET. From our perspective, this time frame is inefficient when you consider how much PET is used daily. As a result, our experiment focuses on increasing the efficiency of the degradation of plastic through the manipulation of PETase. These are just some examples of the types of questions that we must first consider. Once our confidence in our research progresses, we can fully focus on individual procedural steps of our project such as the Western Blot test.</p>
 
 
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<h1 >Abstract </h1>
 
<h1 >Abstract </h1>

Revision as of 00:32, 20 October 2016

The BIG TEMPLATE : RESPONSIVE and FREE

Project Description

Plastic is a ubiquitous material in modern consumer goods. One of the most common plastics used today is polyethylene terephthalate (PET). Chemically, it is a polymer consisting of ethylene glycol and terephthalic acid subunits. With the increasingly short life cycles of consumer products, PET waste is accumulating around the globe at an uncontrollable rate. Moreover, PET is a plastic that does not biodegrade well, and degradation often has to use industrial processes. Given the rapid accumulation of PET and the difficulties associated with biodegradation by ecosystems, it is paramount to find a more efficient method to degrade PET.

Our iGEM project was inspired by the results of a collaborative study between Keio University and Kyoto Institute of Technology, two prestigious Japanese universities. Their research investigated the ability of PETase, an enzyme, to degrade PET plastic using a bacterium known as Ideonella Saikainesis. The success of PETase in the degradation of PET plastic into Polyethylene Terephthalate and Ethylene Glycol sparked our interest in improving the Japanese ecological environment. Thus, the goal of our iGEM project is to create a biobrick incorporating an ideal Anderson promoter to optimize the use of PETase in an E. coli bacteria system. Past iGEM projects centered on plastic degradation, such as those of Turkey 2014, University of Washington 2012 and Darmstadt 2012, also provided us with inspiration on the potential applications for our project, as well as lab protocols.

Our team focused on making a biobrick featuring the PETase enzyme to contribute to the IGEM registry. In addition, we hoped to optimize the initial step of PET breakdown, or depolymerization, so as to expedite the overall process of degradation. To accomplish these goals, we selected several Anderson promoters of different strengths (weak, moderately-strong, and strong) to test for optimizing PETase production. The faster the rate of transcription for the PETase gene, the more PETase enzyme is produced. The greater the volume of enzyme, the faster the rate of depolymerization, since there is more enzyme available to ‘work’ on a plastic sample.

Abstract

In recent years, the production of Polyethylene Terephthalate (PET) has increased rapidly, as a result of low production costs and consumer demands. PET is one of the most common plastic polymers, comprised of repeating monomeric subunits of Terephthalic Acid and Ethylene Glycol. It is frequently used in the manufacture of plastic bottles and clothing fibres, especially in Japan. As avid users of PET-based products, our team decided to research how to optimise the degradation of PET, which takes an average of 450 years to degrade naturally. We were further inspired to pursue this goal with Keio University’s recent discovery of Ideonella sakainesis — a unique bacteria capable of PET degradation. Thus, we have focused our project on the synthesis of an optimal PETase biobrick, which would be included in the iGEM database. Ultimately, our goal is to find an ideal promoter to expedite the production and secretion of PETase.

References

Keio University and Kyoto Institute of Technology. (2016, March 30). Discovery of a Bacterium that Degrades and Assimilates Poly(ethylene terephthalate) could Serve as a Degradation and/or Fermentation Platform for Biological Recycling of PET Waste Products [Press release]. Keio University. Retrieved June 30, 2016, from

https://www.keio.ac.jp/en/press_releases/2016/cb96u90000005501-att/160330_2.pdf


P. (2015). FAQs - Frequently Asked Questions. Retrieved June 30, 2016, from

http://www.petresin.org/faq.asp


Hampson, M. (2016, March 09). Science: Newly Identified Bacteria Break Down Tough Plastic. Retrieved June 30, 2016, from http://www.aaas.org/news/science-newly-identified-bacteria-break-down-tough-plastic
How Long does it take to Decompose - Facts Analysis. (2012, January 24). Retrieved June 30, 2016, from

http://www.hoaxorfact.com/Science/how-long-does-it-take-to-decompose.html


T. (n.d.). Labjournal Metabolism. Retrieved June 30, 2016, from

https://2012.igem.org/Team:TU_Darmstadt/Labjournal/Metabolism


M. (n.d.). Team:METU Turkey project. Retrieved June 30, 2016, from

https://2014.igem.org/Team:METU_Turkey_project


What is PET? (2015). Retrieved June 30, 2016, from

http://www.napcor.com/PET/whatispet.html


Yoshida, S., Hiraga, K., Takehana, T., Taniguchi, I., Yamaji, H., Maeda, Y., . . . Oda, K. (2016). A bacterium that degrades and assimilates poly(ethylene terephthalate) (Doctoral dissertation, Kyoto Institute of Technology, Keio University) [Abstract]. U.S. National Institutes of Health's National Library of Medicine. Retrieved from https://www.ncbi.nlm.nih.gov/pubmed/26965627.