Team:ASIJ Tokyo
The Project
Bacterial cellulose (BC) is a natural biomaterial that is of interest in many fields due to its high purity compared with plant-derived cellulose. We are optimising BC biosynthesis in Gluconacetobacter xylinus, transferring the system to E. coli, and functionalising the material using proteins in order to expand its properties.
BC has seen use in clothing and health foods, but we develop its application to the global issue of water purification. Rising demand for limited freshwater supplies will lead to more than half of the global population suffering severe water stress by 2030. Improved filtration techniques would help relieve this problem.
The porosity of BC and our synthetic attachment of contaminant-specific binding and catabolic proteins make for a flexible, modular water filter. Our manufactured biomaterial would augment water recycling and reclamation on local and industrial scales, helping to alleviate water stress.
Fast Pace PETase
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Project Description
We are surrounded by plastic everyday, specifically PET (Polyethylene terephthalate). Considered as the most common type of polyester, PET is a plastic resin often used for packaging consumer products. We can also find it in our water bottles, plastic toys, and even in the fiber of our clothing. PET is a combination of two monomers known as ethylene glycol and purified terephthalic acid that, when combined, form the polymer PET. The problem with PET is that although it is a convenient material for humans to use, it does not easily biodegrade. As PET accumulates in the ecosystems around the world, it poses detrimental effects on habitats worldwide. That is why it is crucial that we find an efficient way to degrade PET.
Upon conducting our research on plastic degradation, shortly after deciding to focus on this topic, our team came across a joint collaboration research project from Keio University and the Kyoto Institute of Technology, two prestigious universities in Japan. Their paper explained their recent research on PET degradation into terephthalic acid and ethylene glycol using a new bacterium originating from Japan called Ideonella Saikaienis. We were inspired by Ideonella Saikaienis because it was a wonderful PET degrader (containing PETase) from Japan, our current home. Given this, when we realized that the production of PETase was from a uniquely Japanese context, we saw it only apt to center our project around this as it showcases a special (and scientifically recent) feature of the Japanese environment. Consequently, our aim and experimental idea were born: to optimize the use of PETase and hopefully add a new biobrick to the iGEM catalog. Turkey’s 2014 project on PET degradation as well as The University of Washington's 2012 project and Darmstadt's 2012 Project also provided us with inspiration on the potential applications for our project in addition to lab protocol ideas.
We chose to optimize PETase as our goal to make a new type of biobrick available to future iGEM teams. We hope 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 we will be using a western blot test to help us deal with the signal peptides. Currently 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. Before testing such promoters though, we need to isolate 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?” 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 is much too inefficient when you consider how much PET we use daily. As a result, our experiment will focus 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 then fully focus on individual procedural steps of our project such as the Western Blot.
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 <p> <a href="https://www.keio.ac.jp/en/press_releases/2016/cb96u90000005501-att/160330_2.pdfp">https://www.keio.ac.jp/en/press_releases/2016/cb96u90000005501-att/160330_2.pdf</a>
P. (2015). FAQs - Frequently Asked Questions. Retrieved June 30, 2016, from
<a href="http://www.petresin.org/faq.asp"> http://www.petresin.org/faq.asp</a>
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
<a href="http://www.hoaxorfact.com/Science/how-long-does-it-take-to-decompose.html"> http://www.hoaxorfact.com/Science/how-long-does-it-take-to-decompose.html</a>
T. (n.d.). Labjournal Metabolism. Retrieved June 30, 2016, from
<a href="https://2012.igem.org/Team:TU_Darmstadt/Labjournal/Metabolism"> https://2012.igem.org/Team:TU_Darmstadt/Labjournal/Metabolism</a>
M. (n.d.). Team:METU Turkey project. Retrieved June 30, 2016, from
<a href="https://2014.igem.org/Team:METU_Turkey_project"> https://2014.igem.org/Team:METU_Turkey_project</a>
What is PET? (2015). Retrieved June 30, 2016, from
<a href="http://www.napcor.com/PET/whatispet.html"> http://www.napcor.com/PET/whatispet.html</a>
Yoshida, S., Hiraga, K., Takehana, T., Taniguchi, I., Yamaji, H., Maehada, Y., . . . Oda, K. (2016, March 11). Supplementary Materials for a Bacterium that Degrades and Assimilates Poly(ethylene terephthalate). Science, 351, 1196th ser. Retrieved June 30, 2016, from
file:///Users/lisawatanuki/Downloads/PETase supplemental (2).pdf
Yoshida, S., Hiraga, K., Takehana, T., Taniguchi, I., Yamaji, H., Maeda, Y., . . . Oda, K. (2016, March 11). A Bacterium that Degrades and Assimilates Poly(ethylene terephthalate). Science, 351(6278), 1196-1199. doi:10.1126/science.aad6359
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