Human practice, especially comprehensive human practice, is not just a method or step, it provides core support and guidance for our iGEM projects inside and outside the laboratory. This is of decisive value both in the design of the project and in the laboratory operation.

Our project aims to solve the global eutrophication problem by optimizing the microorganism E. coli through genetic engineering and converting waste brown algae into organic fertilizer to achieve environmental protection and resource recovery. In the early stage, we conducted in-depth research on the differences between traditional chemical fertilizers and biochemical fertilizers, interviewed experts and fertilizer factory staff, and learned about the production process and prospects of biochemical fertilizers. Although the project encountered some challenges during the experiment, through expert advice, we have made important progress. At the same time, we conducted interviews with fertilizer stores and fertilizer factories, and planned to conduct science popularization to increase product awareness. Through this project, we are committed to promoting the sustainable development goals and contributing to the ecological balance of the future.

Through the integration and application of human social practice, our projects are more complete, more realistic, and better able to meet the needs of actual society.

As we prepare for the iGEM competition, we review past team efforts to address eutrophication issues "http://2017.igem.org/Team:HokkaidoU Japan" "http://2017.igem.org/Team: OUC-China", aware of the urgency of this issue.

After in-depth research, we found that many previous methods of dealing with eutrophication were simply to remove excess algae and dispose of it. However, we started to wonder: could we find a greener, more sustainable way to deal with this problem? During the course of our research, we discovered algin, a substance ubiquitous in algae that, after proper processing, can be converted into algin oligosaccharides for organic fertilizers.

When our team leader went to PwC to have in-depth discussions with ESG managers, we further understood the concepts of ESG (Environment, Social, Governance) and SDG (Sustainable Development Goals) and discussed the close connection between them. We learned that although chemical degradation methods can effectively degrade algae, its side effects are the production of greenhouse gases and harmful substances, causing further harm to the environment. Although the physical degradation method does not produce harmful substances, it requires a lot of energy and may destroy the beneficial structure of algin. Therefore, we turned to biodegradation methods that are more environmentally friendly and cost-effective.

We chose to use genetic engineering technology to transform the microorganism E. coli so that it has the ability to produce algin lyase. This allows us to efficiently degrade algin into algin oligosaccharides and glucose, thereby obtaining organic fertilizers and other useful compounds, injecting new vitality into ecological balance and sustainable development.

Our team began a long and challenging experiment to continuously optimize the production capacity of the microorganisms and the activity of the enzymes. Over time, we gradually made breakthroughs, successfully achieved efficient degradation of algin, and obtained abundant algin oligosaccharides and glucose resources.

Our innovative approach not only effectively addresses the problem of eutrophication, but also provides a new way of thinking for sustainable development. By converting waste algae into organic fertilizer, we take an important step in environmental protection and resource recycling, making a unique contribution to the ecological balance of the future.

We chose to use genetic engineering technology to transform the microorganism E. coli so that it can produce algin lyase. In this way, we can apply these special enzymes to the degradation process of algin, breaking it down into algin oligosaccharides. This method can not only effectively deal with the problem of eutrophication, but also obtain organic fertilizers and other useful compounds, contributing to the restoration and sustainable development of the ecological environment.

We were honored to invite the team from Nanjing Medical University to share their experience in participating and actively discuss our project at the seminar. We learned more about the shortcomings of the project, which helped us optimize the subsequent projects.

3. Offline training (offline activities)

We all agreed on a common city (Shenzhen) and gathered together. During this period, we designed and shot a project promotional video, created the team's self-media account (such as Douyin, Xiaohongshu, YouTube, ig, WeChat official account), and Published articles and team member introductions. A market research questionnaire for alginate oligosaccharide organic fertilizer was distributed (more than 200 pieces of data have been collected), and companies were searched for interviews both online and offline. Completed the SWOT and stakeholder analysis and content outline of the entrepreneurial plan, determined and confirmed that our project met the three SDG goals, and completed the first version of the hardware design plan and construction.

Our team brainstorms through online meetings and then conducts offline training to improve collaboration capabilities. At the same time, we created a self-media account, conducted market research, and looked for potential partners.

We had intense discussions with 5 teams: RDFE-CHINA, Squirrel-Beijing-1, Beijing Qingmiao International Bilingual School, Beijing Fifth Middle School, and thinker-Shanghai, combining our own team’s projects and targeting the 17 goals of SDG

Source of raw materials:

Traditional fertilizers: usually made through chemical synthesis, mainly including chemicals such as urea, phosphates and potassium salts.

Biofertilizer: Usually made from organic matter, such as animal and plant residues, compost, humic acid and microbial products.

nutrient content:

Conventional fertilizers: Provide a single nutrient such as pure nitrogen, phosphorus or potassium.

Biofertilizer: Contains a variety of microorganisms, organic matter and trace elements to help improve soil health.

Impact on soil and environment:

Traditional chemical fertilizers: Excessive use may lead to soil acidification, reduction of microorganisms and water pollution.

Biofertilizers: Help improve soil structure, increase organic matter content, and reduce negative environmental impacts.

Sustainability:

Conventional fertilizers: Effects are usually short-lived and require frequent applications.

Biofertilizers: The effects may be longer-lasting, helping to improve soil quality over the long term.

Production process:

The production process of traditional chemical fertilizers:

Traditional fertilizers are usually prepared through a process of chemical synthesis, including the use of high temperature, pressure and chemical catalysts.

The production process of biochemical fertilizer:

Biochemical fertilizers are mainly made from organic matter, including raw material collection, mixing and adjustment, compost fermentation, maturation and stabilization, screening and packaging.

Learn about global environmental initiatives, biofertilizer applications and people’s perceptions.

Interview with a Sao Paulo professor from Brazil, focusing on the government’s green economic planning, asking about the impact of chemical fertilizers on soil and the environment, as well as the prospects of biochemical fertilizers.

Brazilian agriculture is mature, but chemical fertilizers have always been expensive.

Chemical fertilizers harm soil diversity and groundwater.

Solutions include adjusting soil pH and developing the organic fertilizer market.

The future development of organic fertilizers needs to reduce costs, improve efficiency and social acceptance.

Understand the process and prospects of biofertilizer production.

Interview staff at a fertilizer factory to learn more about the production process of biofertilizer.

Raw material collection, mixing and conditioning, compost fermentation, maturation and stabilization, screening and packaging.

It has the potential to reduce environmental pollution, improve soil health and increase agricultural production efficiency.

Through this phase of research and interviews we gained an in-depth understanding of the differences between conventional and bio-fertilizers, as well as the prospects and challenges of bio-fertilizer production, as well as the importance of sustainability in the production of algin fertilizer and its contribution to climate warming. Impact on agriculture and the role of biotechnology in agriculture.

At the same time, the consultant also mentioned how alginate oligosaccharide organic fertilizer can protect the soil and ecological environment, as well as ways to ensure the sustainability of the process. Regarding fertilizer selection, consultants noted the importance of choosing the right fertilizer based on specific needs and soil conditions. In addition, it also covers the discussion of the fertilizer market and popular science promotion, as well as the analysis of the advantages and disadvantages of biotechnology.

Business representatives from Thinker-SC and Thinker-Shenzhen teams conducted an offline seminar. At the meeting, we both introduced each other’s product shortcomings and planning plans, and reached a consensus on how to reach cooperation. After several subsequent discussions at online meetings, we basically determined the cooperation model of "seeds combined with fertilizer" sales.

Our team visited the "Six Mu Farm" in Shenzhen and interviewed the farm owner and his employees.

Fertilizer market pain points:

1. High production costs: Organic farms often face higher production costs because they must use expensive organic seeds, fertilizers, and pesticides instead of cheaper chemical products.

2. Low yields: Organic farms often have lower yields due to the inability to use chemical fertilizers and pesticides, which limits their profitability.

3. Limited market: The market share of organic food is relatively small, which makes it difficult for organic farms to find enough buyers, especially in countries like China.

4. Difficulty of certification: Organic farms must go through a rigorous certification process, which is a difficult task for many farms, making it difficult for them to obtain organic certification.

5. High price: Organic food usually has a higher price, which discourages many consumers from purchasing and limits the market potential.

6. Insufficient supply: Organic food production is more complex and organic fertilizers are less efficient, resulting in insufficient supply and making it difficult for consumers to find organic products.

Fertilizer market demand:

1. Reduce costs: In order to reduce production costs, organic farms need more efficient organic fertilizers to reduce dependence on manual labor and improve production efficiency.

2. Expand the market: Since the price of organic food is high and only a few people can buy it, organic farms need to increase production to meet market demand and reduce the price of organic food to attract more consumers.

3. Simplify certification: To help more farms become certified organic, organic farms need to support simplifying the process of organic certification to make it more feasible.

Future prospects:

1. Bright future: The organic food market has a promising future. As people continue to pay more attention to food safety and health, the organic food market share continues to expand. The global organic food market is expected to reach 200 billion US dollars in 2025. China is one of the countries with the fastest growing organic food consumption. The Chinese market is expected to reach 100 billion yuan in 2025.

2. Challenges and opportunities: Although organic farms face many challenges, with the rapid growth of the organic food market, organic farms have the opportunity to reduce costs, increase production and expand market share through innovation, and seize the opportunities of the development of the organic industry.

In the experiment in July, we tried to insert alginate lyase and cellulase genes into engineering bacteria, but found that the activity units of the enzymes did not reach the expected value, and the experiment failed. This means that the efficiency of degrading alginate is much lower than expected.

We later investigated and discussed the function of cellulase with the Squirrel Team, and concluded the benefits of combined hydrolysis, and found that the degradation efficiency would be higher if combined with cellulase.

We sought advice from experts, including doctoral and graduate students Dr. Qin, Mr. Yang and Mr. Wang from Shenzhen University. They pointed out that alginate lyase and cellulase are macromolecular substances and will not travel outside the body of bacteria to degrade polysaccharides outside the body. Large molecules. Therefore, we need to introduce a specific cleavage gene that releases these enzymes. Although they provide these suggestions, they do not specify which specific cleavage gene to use because there are multiple options.

After the first failed experiment, we brainstormed within the team and studied a lot of information about cleavage genes. By reading the literature, we identified some understanding systems that can be used, such as ColM, Lysep, MS2, SRRz, and X174E [1]. We also learned that lytic genes often need to be used in conjunction with an inducible system to release the enzyme when needed. Through investigation, it was found that lac-inducible promoter and ara-inducible promoter can play a role in inducing suicide system function. However, we are still unsure which induction system and lytic gene should be chosen.

In order to understand the induction system and the selection of cleavage genes in more detail, we conducted another interview with Mr. Yang, a graduate student in the field of biology. After exploring multiple potential cleavage proteins and inducible promoters, we ultimately chose to use an arabinose-inducible promoter to initiate transcription and translation of the SRRz cleavage gene. This choice provides flexibility, controllability, and efficiency in experiments and applications, providing a powerful tool for our projects.

Through this phase of adjustments and expert help, we took an important step toward solving the problem and identified the combination of lysis and induction systems that would best suit our project. This will help improve the efficiency of our experiments and make our biodegradation methods more feasible.

1. Bundle sales: We offer bundled packages of seaweed oligosaccharide fertilizer and IAA growth hormone seed packaging, allowing customers to enjoy more discounts.

2. Purchase separately: If customers do not want to purchase a package, they can choose to purchase seaweed oligosaccharide fertilizer or IAA growth hormone seed package separately. Each product's packaging will feature promotional content about the other's product, allowing customers to understand the completeness of organic farming solutions.

3. Additional benefits: For customers who purchase products, we will provide additional benefits, such as free fertilizer when purchasing seeds. Customers who purchase the seaweed oligosaccharide fertilizer package can receive a free IAA growth hormone seed package to try and verify its effectiveness at home.

4. Marketing: We will conduct extensive marketing, including social media promotions, agricultural exhibitions and cooperative projects with agricultural associations. We will emphasize the benefits of organic and natural farming, pollution-free practices, and improvements in crop yield and quality

Future plan:

For the purpose of environmental protection, we plan to sign cooperation agreements with farmers who have been severely eroded in the Loess Plateau area to carry out soil restoration projects. Specific operations include providing high-quality fertilizers at lower than market prices, combined with Thinker-SC’s IAA products for soil restoration and agricultural development. If the project is successfully implemented, it will not only promote local economic development, but also help reduce soil erosion hazards in the Loess Plateau region and society as a whole. This project has an important positive impact on the community.

Our purpose: In order to evaluate the feasibility of our entrepreneurial idea in the market and understand its fit with market needs and prospects, we conducted interviews with fertilizer stores.

What we did: We contacted the owner of a fertilizer store in Shenzhen Nanshan Agricultural Products Wholesale Market:

We took a look at the best-selling fertilizer products sold in the store.

We explored the market share and acceptance of organic fertilizers, and the information we received showed that organic plant fertilizers are still relatively new on the market and will take time to gain wider acceptance.

Store owners also mentioned that marketing for home use can be challenging.

We also discussed the competitiveness of the algin-based fertilizers we produced, especially in terms of price, but the shop owner said that she did not know much about the market situation of seaweed fertilizers, because the seaweed fertilizers of her peers are imported, with higher prices and relatively low sales volume. lower.

She felt our product needed more time to establish its market position.

What we learned: Through this interview, we realized that the needs and expectations of the fertilizer market are complex and diverse, so success does not rely solely on price competitiveness or a single selling point. Organic fertilizers in particular need more time to become widely accepted. Secondly, our fertilizers are more expensive than traditional fertilizers, but lower in cost than similar types of fertilizers. All things considered, given that the manufacturing cost of seaweed fertilizers on the market is relatively high, we consider focusing our target customer groups on organic agricultural product producers who are more willing to pay high prices.

During the conversation, we mentioned that our alginate oligosaccharide fertilizer can play a positive role in reducing climate change and is in line with the climate change goals in the United Nations Sustainable Development Goals (SDGs). The production and use of traditional chemical fertilizers will produce a large amount of greenhouse gases, while algin oligosaccharide fertilizers can reduce greenhouse gas emissions.

Our purpose: We designed hardware equipment aimed at improving the efficiency of biofertilizer production, especially the industrial production of alginate oligosaccharides.

What we did: We designed a device that crushes the seaweed, provides a fermentation environment, and ultimately breaks down the bacteria to release the oligosaccharides. This device is inspired by a meat grinder and is capable of efficiently cutting seaweed into small pieces. The fermentation process uses standard fermentation tanks equipped with precise temperature sensors to ensure temperature control. In addition, we have installed an alarm system to alert users to add ingredients when a specific temperature is reached, thereby destroying bacteria and releasing alginic acid oligosaccharides.

What we learned: Initially, we planned to use the SRRZ gene to control the self-destruction of E. coli, but after discussions with expert MSc Gabriel Saraiva, we decided to use a more traditional high-temperature sterilization method, which has a proven and successful rate High technology. We also recognize that our target customer groups may be more inclined toward increased productivity and mass production, and therefore our hardware may be more suitable for manufacturers.

https://2023.igem.wiki/thinker-shenzhen/hardware

Through this project sharing, we have a deeper understanding of our project and the future development direction.

Through this interview, we have a clearer understanding of the advantages and disadvantages of our project, and it also promotes the development of our business.

Our purpose: During the preliminary fertilizer store interviews, we found that even professional fertilizer store owners did not know much about this new field. With this in mind, we plan to create a public account to conduct relevant science propaganda to increase people’s awareness of new biofertilizers.

What we did: We used our team’s social media to run relevant promotions across the web to increase awareness of our products.

What we learned: Through social media promotion, we have learned how important it is to have adequate publicity to promote new products in the market, and the important role social media plays in people's understanding and influence of products.

[1] Diao, W., Guo, L., Ding, Q. et al. Reprogramming microbial populations using a programmed lysis system to improve chemical production. Nat Commun 12, 6886 (2021). https://doi.org/10.1038/s41467-021-27226-3