Our synthetic biology application project in the field of heavy metal pollution detection involves the following stakeholders: residents, governments, legal, research institutions, industry enterprises, environmental governance companies, etc. We have taken into account the values and considerations of each stakeholder and reflected them in our project execution.
In a city in northeast China, the hometown of one of our team members, a new copper smelting factory was constructed. People there worry that the sewage discharge from the copper smelting plant will pollute the groundwater, thus affecting their health. Some people even worry that the sewage water will be discharged into the nearby river, which will lead to environmental pollution in the whole basin. Therefore, the local people questioned the safety of the factory. Relevant government departments carried out a detailed investigation, invited an authoritative environmental impact assessment agency to conduct a detailed inspection, and issued an environmental assessment report. The report showed that the pollution level of the enterprise met the national standards. However, local residents are still worried about the leakage of wastewater.
We consulted relevant literature and found that industrial wastewater contains heavy metals with significant biological toxicity, such as arsenic, copper, lead, cadmium, nickel, mercury, zinc, and thallium, which have multiple negative effects on human health, including carcinogenicity, mutagenicity, teratogenicity, etc. The pollution of heavy metals is different from that of organic compounds. Some organic compounds can reduce or eliminate their harmfulness through physical, chemical, or biological purification naturally. However, heavy metals are rich and difficult to be degraded in the environment. Some heavy metals in water can be transformed into more toxic metal compounds under the action of microorganisms, such as the methylation of mercury; Heavy metals absorbed by organisms from the environment can be enriched tens of millions of times in larger organisms through the biological amplification of the food chain, and then enter the human body through food, which can accumulate in some organs of the human body to cause chronic poisoning and endanger human health. Minamata disease (mercury pollution) and bone pain disease (cadmium pollution) in Japan, for instance, are typical examples. Therefore, heavy metal pollution should be strictly regulated. The locals are worried that despite the enforcement of governmental regulations, accidents or other factors might still directly influence their health and cause devastating impacts.
Of course, the government has also been taking action to prevent environmental pollution. We have learned that the Ministry of Ecology and Environment of the People's Republic of China has promulgated many laws and regulations and set up many environmental quality monitoring stations to monitor and manage enterprise pollutant emissions. At the same time, enterprises are required to install and use monitoring equipment, regularly detect pollutant emissions, strictly control industrial pollution, and reduce ecological damage. In addition, the Law of the People's Republic of China on the Prevention and Control of Water Pollution clearly stipulates that the state encourages and supports scientific and technological research on the prevention and control of water pollution and the promotion and application of advanced and applicable technologies, and strengthens publicity and education on water environment protection. We are in the middle of a longstanding battle against heavy metal pollution.
Therefore,
First, we contacted experts from the Chinese Research Academy of Environmental Sciences (CRAES). CRAES is the most high-end research institution of environmental science in our country, and it is also an environmental impact assessment agency for enterprises that are obliged to control pollution. They also have a clear understanding of relevant laws and regulations. We want to know:
Experts from CRAES, including Dr. Zhipeng Bai, Dr. Yongjie Wei, Dr. Yan Qian, and Dr. Zhigang Li, held many meetings and communications with us. They were very supportive of our research on environmental pollution and were very interested in the exploration of synthetic biology that we wanted to carry out. They told us:
The main legal basis is as follows:
From the expert interviews and meetings with CRAES, we learned that:
We did not do interviews with enterprises that might cause environmental pollution because it might be a sensitive discussion. Moreover, in China, the government has strict supervision over these enterprises, and the main detection and governance work is undertaken by third-party governance companies. Therefore, industrial enterprises themselves do not know details about environmental governance technologies.
Next, we made an interview outline for the experts from the third-party governance enterprises. We gained a deep understanding of the heavy metal pollution detection work of these enterprises.
1. What business does your enterprise mainly do? Are there many heavy metal pollution detection businesses at present? What are the main customers?
2. What methods are mainly used to detect heavy metal pollution in China?
3. Advantages and disadvantages of these methods? (including but not limited to equipment price, detection price, sensitivity, required time, process complexity, operation difficulty, etc.)
4. What method does your enterprise use? Why choose this method?
5. Has your enterprise used or tried biological/synthetic biotechnology to detect heavy metal pollution? What technologies? What are the advantages and disadvantages of these technologies?
6. What do you think are the main reasons why synthetic biology technology for detecting heavy metal pollution has not been widely used?
7. What technologies are mainly used for heavy metal pollution detection abroad? If it is different from that in China, why is it not used in China?
8. What do you think is needed and can be improved in the current heavy metal pollution detection work?
9. Do you think it is feasible to improve the synthetic biology method for heavy metal pollution detection? If there is such technology, will you and your enterprise use it?
10. Can the method of heavy metal water pollution detection be applied to soil and other environmental detection?
11. Do you have any other suggestions on heavy metal pollution detection, synthetic biology technology, etc?
When we went through the literature, we found a document on the technology and methods of heavy metal pollution detection. We contacted the author of this article, Mr. Chen Wang, and found that he works for SPXFLOW now. He and his boss Mr. Xianbin Yan introduced to us the pollution detection technology ten years ago and the technological development process up to now.
Next, we interviewed experts from Emperor of Cleaning Hi-tech. This is a medium-sized environmental protection enterprise. Their technical and marketing experts, Ms. Shijue Yuan, Mr. Shan Jiang, and Mr. Jianqiang He, enthusiastically helped us. They introduced to us in detail the instruments and equipment currently used in their company and the operation process, and told us that it should be feasible to detect heavy metals via synthetic biological methods. Their company has applied biological methods in practical business, and they are doing experiments on the treatment of heavy metal pollution by engineering bacteria.
Later, we interviewed the detection technology expert of Poten Environment. Ms. Yunhui Fan is very senior and has more than 20 years of detection technology experience. She gave very detailed answers to our interview questions.
Based on the suggestions of all experts, we compared the use of heavy metal detection technology and equipment by different companies and made the following summary:
In the interview with the governance enterprises, we learned that,
Then we consulted synthetic biology experts. They believe that it is possible and feasible to use synthetic biological methods to detect heavy metals.
With the Human Practice above, we determined that,
Therefore, we determined the content of our project: tackling the problem of heavy metal pollution based on synthetic biology.
Through literature research, we found that some people have been trying to detect heavy metals by synthetic biology. However, the sensitivity and accuracy were not ideal. Therefore, we determined the project direction, that is, through experiments, to find a method with better sensitivity and accuracy. At the same time, on the premise of ensuring safety, we developed a convenient, efficient, low-cost and friendly to environment method for detecting heavy metals with synthetic biology.
We called our initial product LABPAT. Taking arsenic pollution detection as an example, it would use standard synthetic biological components BioBricks to synthesize plasmid templates and preserve them. At the same time, the self-developed plasmid template and LABPAT software would be used for actual pollution monitoring as an example to measure its accuracy. Later, a more advanced suicidal module would be added at the same time to prevent the leakage of mutant strains and ensure environmental friendliness and safe discharge. With the LABPAT tool, convenient kits and colorimetric cards could be synthesized, and auxiliary pollution monitoring becomes more popular and convenient.
For this reason, we specially contacted the author of the literature, Prof. Yu Bo, Chinese Academy of Sciences, and asked him to provide the genetic sequence of the suicidal gene.
We invited synthetic biology experts, environmental researchers, and environmental governance companies to help us demonstrate the feasibility of the scheme. We also asked advice from:
After this round of discussion, it was found that:
According to this HP process, after listening to the experts' suggestions, we realized the problems with the first version of the project plan and made modifications. We then adjusted the project from database software to a biosensor. The database was deleted and hardware design, among many other elements, was added to the project.
After determining the project scheme, we determined the experimental scheme. And prepare for the implementation of the experiment in CRAES. Unfortunately, after nearly a month of preliminary preparation, we were unable to enter the high-end national laboratory of the CRAES due to the occurrence of new COVID cases in Chaoyang District of Beijing and the government's requirement that all staff work at home.
As the closure did not end in a short period, we had to look for alternatives to offline experiments in the waiting process. Fortunately, we discovered the concept of cloud lab when we studied last year's iGEM project, so we contacted Ailurus. After discussion, we found that Ailurus’ cloud lab can realize our experimental plan. The advantages of cloud lab include low cost, low technical requirements for the people who proposed the experiment, more data can be obtained in a shorter time, fast iteration, etc. The only problem was that we originally had planned to carry out experiments on arsenic. Ailurus was unable to carry out experiments on arsenic due to limitations of laboratory security. We put safety at a high priority, so we gave up arsenic detection and continued the literature work to find the alternative. We found a paper on a copper-sensitive part and contacted the author Prof. Shoushuai Feng for requesting the gene sequence. Finally, we took copper as the experimental subject.
After the project entered the experimental stage, we communicated with experts from CRAES, the Enterprises, and the synthetic biology experts. They agreed that synthetic biology detection methods for heavy metals pollution maybe a useful attempt and were willing to support our projects. They also looked forward to technological innovation and upgrading. After the initial success of our project in the laboratory, we will further cooperate with CRAES and the governance enterprises to conduct tests in their actual work and continue to improve. We look forward to achieving technological breakthroughs and applying mature synthetic biology heavy metal detection technology to the real world.
To achieve this goal, we continuously referred back to relevant stakeholders in the related fields, making sure that our executive plan met their values and needs.
At the same time, experts in laws and regulations also reminded us that if a new environmental detection technology is to be formally applied in the market, it needs to be evaluated and certified by several research institutions, and can only be formally applied in the market after the formation of nationally recognized implementation specifications.
For this reason, with the support of Duke Kunshan University and under the mentoring of experts from CRAES, we have successfully got the project approval of the 2022 National Innovation and Entrepreneurship Plan for College Students. After iGEM, we will continue to carry out further research and application of our project and apply for industry-standard certification through national procedures and channels.