Team:HK CPU-WFN-WYY/Human Practices

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Human Practice

Overview

Considering the mounting number of microplastic in the ocean, our team determined to design an efficient and cost-effective way to degrade PET(polyethylene terephthalate) in the ocean. In order to prove our project is good and responsible for the world, we asked for opinions from different advisors.

Our Project

World Situation

Nowadays, we can easily find plastic everywhere on our planet. A 2017 study reported that 83% of tap water taken around the world contained plastic pollutants(1).

All of you know that marine plastic pollution is harmful. In order to investigate the impact of it, we have collected opinions from the public and WWF by online questionnaires and email respectively. Survey’s result shows that nearly 100% of 138 participants agree marine plastic pollution destroys ecosystems and disrupts the habitats of wild animals. Plus, WWF said marine plastic pollution is extremely serious as evidence has been found that organisms are killed because of eating plastic and it can also be found in our food and faeces.Thus, plastic may spread unknown bacteria or viruses to humans. Moreover, the degradation of PET product-plastic bottles in the natural environment is very low. It has to take about 450 years for complete degradation(2).

As a result, we need an immediate solution to stop this.

Impact of our project

Our project aims to alleviate plastic pollution around the world. We have designed an efficient way to reduce PET in seawater by using the PET-degrading enzyme, cutinases. It can efficiently degrade PET in a short time, promising 97% in 96 hours. Plus, it can directly digest PET in its final products, ethylene glycol(EG) and terephthalic acid(TPA). Thus, we are designing to incorporate cutinase gene into bacteria that are relatively easy to control in phase 1 and applying it in the ocean in future.This can reduce the amount of microplastic in seawater.


Who will benefit:

  • animals: Our project might save marine animals from being killed by directly ingested plastic or indirectly ingested plastic by preying on bivalves.

  • Seafood consumers: Our project might reduce the amount of microplastics present in seafood. Thus, it might make our food safer and reduce the chance that plastic spreads unknown bacteria and viruses to humans.

  • Environmental communities deal with global warming: As there is a way to decompose PET, the methane and ethylene produced during breaking down of plastic is reduced. So as greenhouse gases are reduced and climate change caused by excess of greenhouse gases is ameliorated.

People who might be opposed:

  • Environmental communities that conserve the ocean: Our project may lower the awareness of plastic pollution of humans. Then it may increase the plastic consumption as people realize that plastic can be directly degraded in a short time.

    Our project may also carry significant risk of creating serious environmental harm. A much better approach is to reduce and avoid plastic pollution at source, through development of a circular economy.

  • Government: As the government might appropriate funds for allowing and providing professional equipment to the research team for further research, money to spend may be one of the concerns for implementation or not.

How do we specialize more than others?

Nowadays, there are various ways to deal with plastic pollution. Including like 3Rs, ocean cleanup and PETase, a PET-degrading enzyme. Therefore, how is our project different from them?

First of all, we aim to degrade microplastic that are already formed in the ocean. Several ways, like 3Rs lifestyle and using wood straw instead of plastic one. They are reducing plastic consumption.However, there are limited ways to deal with the existing microplastic. Thus, we aim to reduce existing PET in the ocean.

Second, cutinase is more efficient than PETase. PETase requires two reactions to digest PET into its final products, but cutinases only require 1 reaction. Thus, cutinases speed up the degradation time. Also, the enzymatic activity of native PETase is low which is approximately 1.7% weight loss in 24hours. But that of cutinase is approximately 25% in 24hours.

Could our project be misused?

Since our project itself is to degrade microplastic in a faster and more efficient rate, people may lower their awareness of plastic pollution after knowing plastic can be broken down in a short time. Then, plastic consumption may increase.

In order to prevent the above consequences, we should promote our project vigorously to the public that our work only deals with the microplastic that was formed in the past. Nowadays, people should reduce the use of plastic in order to conserve the environment.

Fitting Project In Real-World Context

Overview

The ecology, law, effectiveness, desirability and economic factors of our project are the things we most concern about. Thus, in order to prove our work is responsible and good for the world, we have worked with people from diverse communities.The details are shown below.

Ecology

In order to predict and minimize the ecological impact of our project, we discussed with most interested and affected communities.We discussed this with the World Wildlife Foundation(WWF) in email and collected public opinions by analysing their answers in an online questionnaire.There were 138 participants in the survey.

  1. World Wildlife Foundation(WWF)

    2. WWF is an environmentally -friendly organization which has conducted in-depth investigation into marine environment. We ask them about the ethics of releasing GMO bacteria into the ocean and also the biosafety of it.

    • Possible impact: Our precious design which incorporates cutinase gene into a symbiotic bacteria into bivalves , should be considered the risk for the bacteria to spread into the natural ecosystem .For an example, the introduced GM bacteria may pose competition to the natural bacteria ecosystem and infect the natural bivalves. Such infection might affect the biodiversity and ecosystem health of our environment.

    • Our Approaches: The methods we use to minimize the above impact is shown in the Integrated Human Practices Part.

  2. Public Opinions

    • Possible Impact: Our previous project may cause potential safety concerns of bivalves. And it has to go through a lot of clinical practice and can not be lifted at the root of plastic pollution. Plus, its cost may be high.

    • Our Approaches: The methods we use to minimize the above impact is shown in the Integrated Human Practices Part.

Legal

In order to apply current policies and regulations to our project, we discussed this with the Agriculture, Fisheries and Conservation Department, a Hong Kong government department. It is responsible for safeguarding the ocean, ecological integrity and processing of individual GMO approval applications.

  • Government Department- Agriculture, Fisheries and Conservation Department(AFCD)

    • Current policies and regulations: If we are conducting the experiment in the laboratory,we should make sure the GM bacteria is properly disposed (e.g. autoclaved). And thus no GM bacteria will be exposed to the environment.

    • Policies Application: Since we change our project by releasing cutinases in bivalves in future, thus we will inform them about the potential risks of dealing with bivalves in future. The application of above regulation is shown in the Integrated Human Practices Part.

Effectiveness

In order to prove our project benefits the society, we conducted a survey to the public. All of the following results were collected from the online questionnaire which were conducted by our team.

  • Questionnaire Result

    • Public Comment: There are 138 participants in the public questionnaire. Twice as many people who agree that our work is effective as those disagree.Plus, nearly 70% of participants think our work can save marine life, 60% of participants think our work can protect the habitats of wildlife animals. Thus, our work can meet the expectation of the public and likely to bring a great impact to the world.

Viability and Desirability

Our team met with Professor King- Lau Chow in a ZOOM meeting which lasted for almost 1 hour. And he gave us valuable opinions on the feasibility of our project.

  • Meeting With Professor King-Lau Chow

    • Professor King-Lau Chow of Life Science, Chemical and Biological Engineering at the Hong Kong University of Science and Technology. His research focuses on genomics, developmental genetics, evolution and synthetic biology.

    • His opinions: Natural environment may not be suitable for cutinases to work on. As most cutinases’ optimal temperature and pH do not match with that of the ocean. Thus, it may denature the enzymes and lowers efficiency.

    • Our Approaches: The methods we use to minimize the above impact is shown in the Integrated Human Practices Part.

Economy

In order to prove our project is cost-effective, we conduct an in-depth assessment of our work’s cost. The cost of per reaction, solar PET terminator and LB(500ml) will be discussed below.

Integrated Human Practices

Overview

In order to further improve our work, we have met with different advisors online due to the pandemic. They are very willing to give valuable opinions on the ethics,feasibility,laws and technical problems.

Discuss With World Wildlife Foundation(WWF)

  • Their view: The introduced GM bacteria may pose competition to the natural bacteria ecosystem and infect the natural bivalves. Such infection might affect the biodiversity and ecosystem health of our environment.

  • Our Changes:Regarding the unknown consequences of releasing cutinases inside the bivalves. We change our execution by using cutinases in a controlled and closed environment in phase 1, instead of incorporating GM bacteria symbiotic to bivalves. The previous design will be done in the future when risks and consequences are evaluated concisely.

Discuss With Professor King-Lau Chow

  • His view:Natural environment may not be suitable for cutinases to work on. As most cutinases’ optimal temperature and pH do not match with that of the ocean. Thus, it may denature the enzymes and lowers efficiency.

  • Our Changes: Do it in an enclosed environment where temperature and pH are controlled in phase 1. And we compare the temperature and pH of 6 more cutinases, in order to find the most suitable one. Furthermore, we design a solar degradation device in order to let cutinases work in a more favourable place and increase the enzymatic activity. Since the temperature is near the glass transition temperature of PET, it also increases the degradation rate.

  • Detail of solar degradation device: It is called solar PET terminator, which composed of 5 main parts: solar water heater, degradation tank with the immobilized cutinase coated on beads, a battery, a heater and peltier modules.

  • Its mechanism: The solar water heater absorbs sunlight and turns it into heat energy to heat up the water.The heated water will flow to the degradation tank.The degradation tank contains beads with immobilized cutinases, which degrades PET in water.

  • Work In Special Conditions: When the water temperature is too high, an electric solenoid valve will open and let the water flow to the peltier modules. On cloudy days or when water temperature is too low in cold winter, the heater will draw electricity from the battery to heat up the water to the optimum temperature for cutinases.

Discuss With Local Government Department-Agriculture, Fisheries and Conservation Department

  • Their view: If we are conducting the experiment in the laboratory,we should make sure the GM bacteria is properly disposed (e.g. autoclaved). And thus no GM bacteria will be exposed to the environment. Moreover, we should take a look on the local policies of releasing GM organisms.

  • Our changes: We deeply look for the local policies and check if our project is legal. For example, we look for the Genetically Modified Organisms (Control of Release) Ordinance and understand we need to apply the documentation requirement and receive approval of our GM bacteria before releasing it to the natural environment.

First Meeting With A Technician

In order to produce a better presentation video, we have several meetings with a technician- Mr. Lau. We asked him the opinions of our promotion video and presentation video at the ZOOM meeting due to the pandemic.

  • His view: It used to be very time-consuming to add subtitles to the promotion video. And it was also very hard work.

  • Our Changes: We create a subtitle generator to speed up the process of adding subtitles.

  • Details of subtitle generator: Users can directly exchange the video format VTT and SRT, which are the common video format of iGEM2020. It would speed up the process of changing video format.

    Moreover, as long as users type the subtitles in a specific format, the program will then automatically combine the subtitles in the film. It saves editing time as users do not need to subtitle each sentence and thus avoid being exhausted. Plus, the users can adjust the playback speed and thus reduce the time it takes to make a film. In addition, this generator can hold video for up to 99 hours. So, users do not have to worry about it not being able to accommodate their video.

Second Meeting With The Technician

We met with Mr. Lau at the ZOOM meeting again, but this time we showed our subtitle generator to Mr. Lau.

  • His view: The subtitle generator is very helpful and user-friendly. Since users can easily exchange the video format VTT and SRT, which are the common video format of iGEM2020. Moreover, as long as we type the subtitles in a specific format, the program will then automatically combine the subtitles in the film. Thus it saves editing time and benefits other users.

Education And Public Engagement

Overview

Apart from working on our project, we have shared our ideas and explored more angles of synthetic biology with the public. In the meantime, it spreads the influence of our project.

Post Mini Talk

The mini talk was held by City University of Hong Kong, the Hong Kong University of Science and Technology and our team at the ZOOM meeting. The participants were mainly high school students. The total participants were around 167 people. We introduced Synthetic Biology, career pathway related to biotechnology and examples of local companies.

  • Result: We have conducted a survey about the talk to 108 participants. The result is collected from the online questionnaire. Results show that more than 85% of participants agree that the talk is beneficial to them. For detail, they agree that the talk enhances their understanding on the job content related to Synthetic Biology and the career pathways of Synthetic Biology.

Online Video Learning

Problem of microplastic has become more serious. Thus we want to spread this message and let more people be aware of marine plastic pollution. Therefore, we have provided two learning materials for high school students and test their understanding of them.Their feedback of the video was collected from an online form.

  • Result: According to our survey, more than 87% students gain an insight in microplastic, raise awareness on plastic pollution and more willing to reduce the use of plastic Moreover, some students relies that the video let them rethink how they can reduce the use of plastic everyday.

Collaboration

HK iGEM Team Symposium

All Hong Kong iGEM Teams and various advisors participated in this symposium. Each team was required to present their works and Q&A sessions were provided for advisors to give several opinions to the projects.

    Opinions:

  1. The cost of our project may be higher than that of the traditional way to break down PET.

  2. Our enzyme may be easily inhibited.

    Our change:

  1. We make an in-depth assessment about the cost of our project. Eg: the cost of the production of GM bacteria is $130 USD despite the cost of equipment

  2. We also make an in-depth research of inhibitors for each cutinase. And we found that cutinases are not easily inhibited. On the contrary, there are many mediators to enhance their activity and thermal stability. For example, Ca2+, Mg2+ and lipase CalB.

Online Biosafety and Biosecurity Seminar

Our team participated in an online talk with a professor from The Open University of Hong Kong, taught basic knowledge of biosafety and biosecurity.

  • Our change: All team members are more concerned about the biosafety of our project.For example, we have searched the pathology of several cutinases.

Reference

(1) https://www.theguardian.com/environment/2017/sep/06/plastic-fibres-found-tap-water-around-world-study-reveals

(2) "https://www.wwf.org.au/news/blogs/the-lifecycle-of-plastics#gs.j41t0e