Human Practices is the study of how your work affects the world, and how the world affects your work." — Peter Carr
We focus not only on synthetic biology, but also on whether our research is beneficial to the world. Therefore, HP is also an important part of our project, which even is the general ideological guidance of our project. We believe that with more people understanding and engaging in our project, we can carry it out better and make it more beneficial to the world. Although COVID-19 has partly hindered our plans to go out for practice and research, we don’t stop our work and carry out many activities with the help of Internet tools.
To this end, we designed a questionnaire to listen to the public's wishes; communicated with experts to improve our project from different directions: consulted the government to understand the treatment of plastic pollution; visited factories to learn the practical application of PET. What’s more, we launched a series of online courses to let more people know us further; we helped poverty alleviation and sown the seeds of hope.
Before the start of our project, we were at a loss as to the value of our project and what needs to be improved. And we did not communicated with more people, we didn't know whether our project can be accepted and supported by most people.
Therefore, to understand the public's awareness of plastic and plastic pollution, to know the public's views and suggestions on our project, and to better improve our project, we produced and released a questionnaire. After four months of collection, we collected a total of 748 replies from home and abroad, we were fortunate enough to be able to spread this survey to international citizens. We received responses from France,Pakistan,United States,Germany, Britain,Tanzania and Netherlands.
At the same time, we also received many questions, concerns and suggestions, which not only provided practical ideas for our later introducing biodegradation and our project to the public, but also make us pay more attention to biosafety issues in next year's experiments.
From the responses , we know that nearly 75%of responses are comprised of those aged 18 - 25 and 41 - 65 ( Fig. 1). It is relieving to know that a large majority has received high school-level education at the very least (Fig. 2), although only 20% are of biology or chemistry backgrounds ( Fig.3). Therefore, our survey is rational, reliable and representative.
Views on plastic pollution
Firstly, we wanted to assess the sample population’s general view of plastic pollution. We set up three different questions to reflect the public's perception of plastic pollution from three perspectives.
We are surprised to find that more than 87% of the people think that the current plastic pollution is relatively serious (Fig. 4). Nearly 50% of the people think that their lives are more seriously troubled by plastic pollution (Fig. 5). Nearly 85% of the people do not know the harm of plastic pollution (Fig. 6). From these responses, We learn that the plastic pollution in the world is a serious problem. The plastic pollution has seriously affected our life, and the problem of plastic pollution needs to be solved urgently. These result strengthen our determination to make high-efficiency enzyme for degradation of PET. At the same time, we also see that although people think that the problem of plastic pollution is very serious, many people still do not understand the harm of it, which makes us realize that we have the responsibility to strengthen the publicity of plastic pollution hazards, so we opened a social platform to increase publicity.( See Publicity)
Use and recovery of PET
Last, we would like to know about the use and recycling of the refractory PET bottle.
We find that more than 75% of people use PET bottles every week (Fig. 7), More than 75% of people put PET bottles in a designated place (Fig. 8), which is very gratifying. However, it is sad that only 35% of the people know where PET can be recycled locally, which indicates that the PET recycling system has not received enough attention. Even there are no institutions for recycling PET in some places, which indicates that the PET recycling system has not been widely used. To understand the general content of PET recycling system, we contacted a company specialized in recycling PET，Incom Recycle Co.,Ltd. （ See Enterprise Visits）
In the process of our project, we were full of confidence but suffered a great blow. We consulted a lot of information and conducted in-depth discussions, but we were still confused by some problems and we couldn't find a way to solve them.
Before the experimental results are completed, we must ensure that our theoretical knowledge is correct and convincing, but our mastery of relevant professional knowledge is not enough to enable us to draw correct conclusions. However, when we consulted the information, we found many experts, who were more thorough in their research. Therefore，at the early stage of entire project, we decide to listen to related experts’ advice first. So, to help further verification, improve our ideas and increase the credibility of our results, we visited several experts in different fields.
Dr.Kohei Oda is the first discoverer of PETase. At the beginning of our project, we didn't know many details of PETase. Therefore, we planned to communicate with the discoverer of PETase, Dr.Kohei Oda. Since 2016, he has become a Fellow of the Japan Society for Bioscience, Biotechnology, and Agrochemistry. He discoveried novel families of peptidases, sedolisins and eqolisins. His ongoing interests are also microbial hydrolytic enzymes involved in degradation of poly (ethylene terephthalate) (PET).
We sent an email to the Kohei Oda team who discovered PETase to share our team’s efforts of improving the thermal stability of PETase. We put forward many problems and doubts and ask suggestions for our project. They shared with us the latest developments related to our research. According to their answers, we optimized and selected the structure of PETase. Finally, we chose the protein structure with PDB number of 5xjh for our follow-up study. We express our heartfelt thanks for this.
Dr. Nick Pace
Dr. Nick Pace is an expert in biochemistry, who was a visiting professor at Osaka University in the Institute for Protein. He has a lot of research in the field of protein. After reading his article, we were greatly inspired, but at the same time, we encountered a lot of puzzles.
Prof. Arieh Ben-Naim
Prof. Arieh Ben-Naim is a professor in the Department of Physical Chemistry at The Hebrew University in Jerusalem. He has a lot of research in the field of Hydrophobic interaction.
When we studyed hydrophobic interactions, we encountered some problems. We could not explain the essence of this interaction from the atomic structure level. Therefore, we emailed Dr. Nick Pace and Prof. Arieh Ben-Naim. They shared their opinions enthusiastically and provided us with the latest research papers. With the help of Dr. Nick Pace, we understood the problems that puzzled us for a long time. After solving these problems, we have designed more reasonable and efficient hydrophobic sites, such as Q127L. These very good hydrophobic sites were eventually used in our PET-CRUSHER. With the help of Prof. Arieh Ben-Naim, we solved more questions about hydrophobic interactions. After solving these problems, we designed more reasonable and efficient hydrophobic sites. According to Dr. Nick Pace's answer, we found many other hydrophobic sites. Finally, we combined Q127L,S214H and K259F, and the ΔTm of this conjugate has increased by 10.90632. This is a very good result, which makes us very happy.
Mr. Zhu Cheng
Molecular dynamics simulation (MD)is one of the important parts of our project this year. When we encountered some difficult problems in the process of MD, we consulted Mr. Zhu Cheng who is professional in bioinformatics. He provided a lot of guidance for our MD work. After a whole year learning ,now we have grasped the principles and operating methods of MD, even are able to evaluate the results reasonably which is great significance to the promotion and completion of our project.
Dr. Yao Mingdong
The design of protein hydrophobic interaction is one of the essential parts of our project. During the whole season, we kept asking various experts for the principle of protein hydrophobic interaction. Dr. Yao Mingdong, an expert in the field of hydrophobicity, assists us to analyze comprehensively from different sides and levels in protein designing involving hydrophobic part. What’s more, he also provided some suggestions on improving the thermal stability of protein. These suggestions inspired our ideas about tightening some of hydrophobic cores founded by residues located in some loose loops in PETase which really helps for the further improvement of our project.
The performance of our personal computer is far from the requirement to run MD which may cost about 20 days to process a file. So we decided to install GROMACS on the college's high-performance computers. However, as a non-computer major student, we encountered many problems in the process of installing a strange Linux system. Therefore, we consulted Guan Fenghui, who is familiar with the Linux system. With her help, she not only installed the software, but also learned how to submit MD tasks remotely. In addition, Ms. Guan also provided us with Rosetta course to help us learn and use more bioinformatics software smoothly.
Tianjin Development and Reform Commission
To understand the methods in dealing with plastic pollution of government, we first consulted the Tianjin Development and Reform Commission. During the online interview, the staff warmly communicated with us and provided a notice on dealing with plastic pollution jointly issued by nine departments of the government in July. What’s more, they further provided us with the newly released Two Notices on Improvement of Plastic Pollution in Tianjin. When it came to the future planning, they mentioned that the three documents issued were currently an outline for guidance, and many specific rules would be issued in succession. Recently, the government has paid much attention to the environment issue. According to these notices and the communication with the staff, we were impressed by the determination of the government to solve environmental pollution, which deeply strengthened our confidence in this year's project.
Tianjin Ecological and Environment Bureau
After that, we consulted Tianjin Ecological and Environment Bureau, hoping to find out the solution to handle the waste of PET products. The staff told us that China is increasing the recycling process of PET bottles by supporting specialized enterprises and organizations. In addition, there are relevant personnel who specially sort out the PET bottles for special treatment. However, physical and chemical methods are still widely used to deal with PET products, which may cause environmental pollution. Credit to the explanation of the staff, we learned that the government has great determination to solve environmental problems caused by the accumulation of PET through encouraging enterprises to participate in environmental protection. Last but not the least, they were extremely interested in new plastic-waste solutions. All in all, according to the communication and online interview with government departments, we are greatly encouraged and strongly feel that our project may play an important role in environmental improvement, which makes us determine to achieve our project better confidently.
Considering our project is about biodegradation of PET, we’d like to know whether our method is worth putting into industrial production. So we chose to visit the relevant factories to deepen our understanding of the project.
Tianjin Industrial Zijiang Packaging Co., Ltd.
Following the premise of meeting epidemic prevention requirements, we visited Tianjin Industrial Zijiang Packaging Co., Ltd. and gasped a series of data and information about the production of plastic bottles from the company. We were surprised by the huge quantity of 170,000,000 coca cola bottles they produced per year. What’s more, we knew that the crystallinity of their plastic bottles was 35%, whose degradation is the target problem of our project. They agreed that the current plastic pollution and the recycling progress were limited. So we introduced our biodegradation method and project to the staff. They thought it would be a good way to solve the plastic pollution. They appreciated that the method could be vigorously promoted if it was effective.
COFCO Coca-Cola Beverages (Tianjin) Ltd.
After learning about the production of PET, we applied to visit some other companies which use PET. Our first choice is COFCO Coca-Cola Beverages (Tianjin) Ltd. The staff showed us the Coca-Cola production line with great enthusiasm. We learned that Coca Cola Company launched the "world without waste” campaign. The slogan was also committed to promoting the closed-loop economy of PET packaging products which jointly developed recyclable plastics (RPET) with some foreign technology companies. Nowadays, all Ciel drinking water (sold in Mexico) of Coca Cola has been packaged with RPET.
However, due to the traditional physicochemical degradation method, there’re still some problems such as high cost and large mechanical loss. The RPET products are limited since the price is high, so it is difficult to popularize in a wide range. We also introduced the biodegradation method to the staff of Coca Cola Company, and explained in detail the specificity of PETase for high crystallinity PET and our efforts to improve its thermal stability which would promote the industrial application of the enzyme. We even showed that enzymatic hydrolysis would further promote the realization of closed-loop economy of PET, which had aroused great interest of the staff. They were very interested in our project and were willing to provide bottle-grade PET materials in our experiments in the coming year.
Incom Recycle Co.,Ltd.
Last but not the least, to understand the market application prospect of our modified PETase. We consulted the company specializing in PET recycling, Incom Recycle Co.,Ltd. Their current solution of recycling PET is still based on the traditional physical and chemical methods. According to the staff, the physical method they currently use can regenerate PET products for two different purposes, one is downgraded to produce polyester clothing, and the other is to produce PET bottles. However, the crystallinity and strength of the regenerated PET bottles will be greatly reduced. It can only be used as a mineral water bottle instead of a carbonated beverage bottle. This is a great loss.
Therefore, we introduced our project this year to them and stressed that our biodegradation method would not reduce the crystallinity and strength of PET products.
After listening to our project introduction, they said that their company was willing to cooperate with us in depth and try our biodegradation method if it is finally proved to be feasible. In addition, they proposed that the biodegradation method was not only great significant to their company, but also to the whole PET recycling industry. Our efforts to modified PET degradation enzyme can significantly save their costs and increase their recovery efficiency. They were looking forward to our achievements.