Sustainable

Sustainable development is one of the most crucial consideration in our project. We aim to turn current kitchen waste which can contaminate the environment and is hard to be degraded into valuable things that can be beneficial to people who are in need of health care drugs. We aim to build a project that contributes to not only a cleaner environment but also a healthier community. Although our project has numerous sustainable development, we lay emphasis on six out of seven goals-- #3 Good Health and Well-Being, #4 Quality Education, #6 Clean Water And Sanitation, #9 Industry Innovation and Infrastructure, #12 Responsible Consumption and Production, and #17 Partnerships For The Goals.

#3 Good Health and Well-Being

Our project aims to undermine the current significant global health issue that the number of people suffering from the health risk of hypertension, hyperglycemia and hyperglycemia rising without any sign to become flat. Among that, China is the country with the most serious problem. In China, nearly 350 million people are suffering from the health risk of hypertension, hyperglycemia and hyperglycemia[1]. Hence, we hope our end product can help people globallyl to deal with and ease those health risks.

In the project, we produce a health care sugar--chitosan oligosaccharides, which have excellent biological activities, including antioxidants, lowering blood pressure, lowering blood lipid, and anti-inflammatory[2].

After studying the existing technologies, we find that those technologies generally have problems such as low efficiency and the inability to control the output. We finally discovered and used GM technology, planning to combine the plasmid fragment of chitosanase with the plasmid of Escherichia coli to form a new complete plasmid and express it in Escherichia coli.

During the production process, in the laboratory, we have done several tries, including combining a new plasmid that carries the chitosanase gene and propagating it in E. coli Dh5alpha and introducing the ice crystal nucleoprotein gene. The second try is successful. The second time, we still used transgenic technology to extract the chitosanase gene, but in advance, we combined the chitosanase gene with the N-terminal gene of ice crystal nuclear protein into a plasmid. When the chitosanase gene fragment is combined with the Escherichia coli plasmid, the N-terminal gene of ice crystal nuclear protein can convert the intracellular display chitosanase gene into extracellular, that is, the cell surface display technology is used. In the end, we successfully, and efficiently produce our product- chitosan oligosaccharide. Additionally, for sustainable development in good health and well-being, we interview the old, the target user of our product, and find that most elderly people do not understand our product, but they do accept the chitosan oligosaccharide and transgenesis technique. However, they’d like to follow the suggestions from doctors and veterinarians.

As a result, in the future, our product is promised to help people to reduce health risks.

#4 Quality Education

Our team devotes itself to educating the public about synthetic biology. To meet this goal, we work on public account promotion and public lectures

1. Public Account
Our target group is students who are interested in the iGEM competition or synthetic biology and parents who need to know about the competition. Therefore, we set up an iGEM official account, where we can publish content. So far, our team has released four contents, which are an introduction to the iGEM competition, the 7.17 Communication Conference, an introduction to our project, and an explanation of basic synthetic biology. We hope to share our experience with others by publishing official account, and the follow-up feedback we received is that each article has been read by a certain amount, which proves the effectiveness of our educational activities.

2. Public Lectures
We have held three lectures on synthetic biology and AP biology in Beijing 101 Middle School and Qingdao No. 9 Middle School for many students who are interested in genetic engineering.

2.1 The first lecture
In the first lecture, we mainly explained:
1. The cell surface demonstration technology used in this experiment -- adding ice crystal nuclein to produce chitosaccharides more efficiently;
2. The quaternary structure of protein and the dehydration and condensation process of amino acids;
3. The characteristics of carbohydrates, lipids, proteins, and nucleic acids;
4. PCR principle and machine operation method;
5. Steps of plasmid extraction and the role of plasmid construction.
After class, many students actively asked questions and discussed more synthetic biology knowledge with us. Finally, we received feedback that this activity helped Grade 10 students a lot, which made them have a preliminary understanding of the iGEM competition and determined their future learning direction.

2.2 The Second lecture
In the second class, we adopted the online conference to enable more students to participate in it. In consideration of the time conflict, we recorded the class for everyone to watch repeatedly. This time we will talk about:
1. The structure and properties of water molecules;
2. Bonds between different kinds of polymers;
3. Benefits of chito-oligosaccharide and its production materials;
4. Application fields of genetic engineering.
Since we have changed the way of teaching, more students are actively asking questions in the comments section. Because this lecture we explained more details and principles based on the first lecture, Grade 10 students feedback that it is also very helpful for their campus biology courses.

2.3 The Third lecture
The third class was held in Qingdao No. 9 Middle School. Our teaching content was very rich, including:
1. The history of synthetic biology;
2. Concept proposal;
3. Research methods and tools;
4. Research direction.
After the lecture, our team members got support and positive feedback from students, and quite a few students who attended the lecture had a relatively complete understanding of synthetic biology.

We have done a lot to promote the concept of synthetic biology and our product. We hope that through our actions, the public can learn more about synthetic biology.

#6 Clean Water And Sanitation

Recently, challenges are mounting for one of Europe's largest fisheries for crab and lobster amid continued wash-ups of dead crustaceans in northeast England that are compounded by a new phenomenon -- 'dissolving' lobster eggs.

Fishermen in the area of northeast England found this phenomenon and kept records. In a recent video, they tweeted, floating substances can be seen on the water's surface. They described Robin Hood's Bay as a “foul eggy smell yesterday." Additionally, the catches of brown crab remain significantly lower off the coast of Redcar and Hartlepool, the epicenter of an ecological crisis that has been ongoing for a year and one of Europe's most productive fisheries for lobsters and crab.[3]

This pollution critically threatens the life of fishermen and people near northeast England since it causes marine animals to die in numerous amounts and accumulate near water, which leads to the breed of bacteria and thus can contaminate the using water of people. However, in our project, we recycle the overwhelmed, detrimental crab shell as raw material to produce health care product-- chitosan oligosaccharide.

Therefore, through this project, we can solve the problem of excessive crab shells that pollute the using water, and thus provide a clean water environment for people.

#9 Industry innovation and Infrastructure

In the project, we produce an end substance- chitosan oligosaccharides. At present, the main ways of obtaining chitosan oligosaccharides are enzymatic, chemical, and physical methods, which both have disadvantages. The disadvantages of single enzymatic hydrolysis are that the separation and purification of the enzyme are complicated, the enzyme is easily inactivated, and its hydrolysis efficiency is very low. Physical hydrolysis can make the product impure. The chemical method leads to pollution of the environment and products being more mixed, and the chitosan oligosaccharides with a certain degree of polymerization cannot be obtained. On the contrary, a novel method was proposed in our project to utilize Escherichia Coli cell-displayed chitosanase (CHI-1) to degrade the co-fermentation of Bacillus subtilised Acetobacter sp. from shrimp shell waste. After examination, we find that under the best optimized co-fermentation condition (5 g/L yeast extract, 10 g/L K2HPO4, 6% ethanol, 50 g/L glucose), the deproteinization and desalination efficiencies of shrimp shell waste were 94 % and 92%, the yield of chitin was 18%. The surface structure characteristic and functional groups of prepared chitin and chitosan were determined by scanning electron microscopy and Fourier transform infrared spectroscopy, indicating that chitin and chitosan were successfully extracted from shrimp shell waste by co-fermentation of Bacillus subtilis and Acetobacter sp. We also construct the cell surface-displayed chitosanase (E. coli BL21-pET23b(+)-NICHI) in the experiment. Compared with crude chitosanase solution (crude CHI-1), E. coli BL21-pET23b(+)-NICHI showed better hydrolysis ability, E. coli BL21-pET23b(+)- NICHI maintained 81% of its initial enzymatic activity after 40 days left under room temperature. The crude CHI-1 enzyme solution reacted with the prepared chitosan for one day, the crude CHI-1 enzyme solution was almost inactivated. However, E. coli BL21-pET23b(+)-NICHI still maintained a good hydrolysis ability after reacting with the prepared chitosan for seven days, and the yield of chitosan oligosaccharides obtained by its hydrolysis was 41% after 7 days. The chitosan oligosaccharides produced by hydrolysis were identified as chitobiose, chitotriose, maltotetraose, chitopentaose, and chitohexaose by triple quadrupole LC-MS. It can be seen that the technology of displaying chitosanase on the cell surface increases the stability and hydrolysis ability of chitosanase, which can be effectively used in the preparation industry of chitooligosaccharide, improve the utilization value of wastes such as shrimp shells, and reduce environmental pollution[4].

This innovative method not only can be beneficial to the environment, since it uses shell waste but also can be more efficient and time-saving, compared to those three methods previously.

#12 Responsible Consumption and Production

In our project, we use a new method to produce our product- chitosan oligosaccharide, therefore, it is our responsibility to consider sustainable consumption and production. Our raw material- chitin is widespread in nature. It exists mainly in the insect carapace, shells of crustaceans, and cell walls of certain fungi. When we eat shrimp and crab every day, we produce shrimp and crab shell waste which contains a lot of chitin. Chitosan oligosaccharides with better water solubility, more convenience to use, and better biological activity can be obtained by further hydrolysis of chitin[4]. To be more specific, by using shell kitchen waste, we can produce Chitosan oligosaccharides.

By carefully considering the pollution and consumption from nature, we came to the conclusion that our project does not damage or contaminate the environment, by contrast, we are actually environmental-friendly since we help to recycle shell waste that can hardly be crushed or burned.

To draw a conclusion, we have responsibly been concerned about the sustainability of consumption and production.

#17 Partnerships For The Goals

A good project means that we need to have a broad influence and collaboration with others. SDG17 suggests that our team needs cross-sector and cross-country collaboration for pursuing our goals[5]. As to meet this criterion, we find Team LZU-hS-China-C and Team BFSU-ICUnited who also use carrier protein and cell surface display technology as partners. We collaborate in many ways. In the first place, we, three teams together, find six papers related to INP each. Consequently, according to those six articles, three teams select two of them respectively and write a report, which was shared in the joint conference of the three teams, that expands the knowledge reserve of the three teams on INP and provides a reference direction for the use of cell surface display technology of each team.

In addition, each of the three teams has launched its own Wechat account. In their respective public accounts, three teams introduce their teams and project and send them to the cooperative group. The person in charge of each team forwards the public accounts of the other two teams to help each other promote their project.

Thirdly, After discussion and confirmation of the three team leaders, it was decided to establish an educational alliance of three teams, and each team will carry out synthetic biology education and general knowledge publicity in the name of the alliance. Each team will conduct a synthetic biology education in the name of the alliance, and the process record will be forwarded to the group chat of the three teams.

In conclusion, we have cooperated to achieve our goals together for a shared future.

References

[1] (2022). Retrieved 29 September 2022, from https://view.inews.qq.com/wxn2/20220407V02QQZ00

[2] Kaur S., Dhillon G. S. Recent trends in biological extraction of chitin from marine shell wastes: a review [J]. Crit Rev Biotechnol, 2015, 35(1): 44-61

[3] Harkell, L. (2022). Dissolving lobster eggs, crab die-offs; evidence mounts of pollution impact in NE England... Retrieved 5 October 2022, from   https://www.undercurrentnews.com/2022/10/03/dissolving-lobster-eggs-crab-die-offs-evidence-mounts-of-pollution-impact-in-northeast-england

[4] CHITOOLIGOSACCHARIDES PRODUCTION FROM SHRIMP CHAFF IN CHITOSANASE CELL SURFACE DISPLAY SYSTEM. (2022) (2nd ed., pp. 1-40). Beijing, China.

[5] Wikipedia Contributors. Sustainable Development Goal 17. Wikipedia. Published September 12, 2022. Accessed September 29,2022. https://en.wikipedia.org/wiki/Sustainable_Development_Goal_17