Team:ASIJ Tokyo

Project Overview

What is PET?

PET is a combination of two monomers known as ethylene glycol and purified terephthalic acid that when combined form a polymer of polyethylene terephthalate. Considered as the most common type of polyester, PET is a plastic resin often used for packaging consumer products. The problem with PET is that, though it is a convenient material for humans to use, it is also very hard to biodegrade. As PET accumulates in the ecosystems around the world, it poses detrimental effects on habitats worldwide.

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

FAQs - Frequently Asked Questions. FAQs. PET Resin Association, n.d. Web. 07 June 2016.

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

"What Is PET?" NAPCOR. NAPCOR, n.d. Web. 07 June 2016.

http://www.aaas.org/news/science-newly-identified-bacteria-break-down-tough-plastic

“Newly Identified Bacteria Break Down Tough Plastic” AAAS.AAAS, n.d. Web. 25 June 2016

Our project focuses on using the enzyme PETase for breaking down PET hoping to increase the efficiency with which the breakdown occurs. PETase uses the process of hydrolysis to breakdown PET, enabling PET as an energy source for the PETase producing strain 201-F6, and its ability to exist in natural surroundings.

PET (Polyethylene Terephthalate) is a polymeric plastic resin comprised of monomers of ethylene glycol and terephthalic acid.

Inspirations

Using Experimental aids and inspiration from the joint collaboration research project from Keio University and the Kyoto Institute of Technology, and their research on PET degradation to Terephthalic acid and Ethylene glycol using the bacterium Ideonella Saikaiensis. Experimental description of the experiment:

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

Our Plan and Goal

Combining research methods from previously done experiments with what we learn from our own experiments (though we haven’t done many yet!). Right now we want to isolate the first step of our procedure and increase its efficiency, before delving in any deeper. We do have a goal in mind: to add in a new biobrick using PETase to increase efficiency and usability in the plastic degradation process.

However, we’d like to also think about the practicality of our experiment in the real world. Questions we may ask ourselves during our experimental procedures will help us seek solutions useful if applied in the real world. Such questions include: “how long will the degradation take?” and “will the byproducts of the degradation be harmful in any way to the environment or to humans?”. In regards to the first question, to our current knowledge, 6 weeks is the record for the biodegradation of PET. In our eyes, this is much too inefficient. As a result, our experiment will focus on increasing 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 research progresses, we can then focus on individual procedural steps, such as western blot tests, of our project.

The End Products

We’d like to synthesize a plasmid coded to efficiently secrete PETase. The results we hope to see include a larger amount of PET degraded in a shorter amount of time. Hopefully, we can break the record of 6 weeks. This is our main objective in terms of the lab. Theoretically, our project extends to the production of a feedback loop in which we take advantage of tryptophan to create a trp operon system through a series of combined intermediate enzymes. If ever this theory is substantiated, we hope to see bacteria use PET as a food source in which PETase will biodegrade PET when it needs to. In a real world situation, if PET is accumulated in one area where our genetically engineered bacteria have habituated, then we predict that we can leave the PET there to be degraded by the bacteria and basically forget about it. It will disappear eventually. This would requires minimal human and industrial intervention of the disposal of PET. We hope our project will be able to reduce the impact of PET as a pollutant on the environment.