Product research questionnaire

Questionnaire promotional poster (QR code has been invalidated)

Questionnaire analysis

Based on the analysis of questionnaire data, we obtained a total of 690 valid data questionnaires. Among them, the largest proportion is under 18 years old and between 41 and 50 years old. The highest proportion of income is below 3000 RMB. The majority of people who have a basic understanding of sugar substitutes account for approximately 46.81%. The main reason why the public chooses sugar substitutes is still focused on maintaining their figure or losing weight, which accounts for 59.78%. The understanding of sugar substitutes is still relatively shallow. Other reasons are diabetes or other diseases, but they are less than the previous one. Since people believe that sugar substitutes can help maintain their figure or lose weight, they hope that sugar substitutes can be a product that assists in controlling sucrose intake. These products are not completely absent from the current market, such as sugar-free Coca-Cola and Pepsi Cola, which have high sales and bring high profits and also help control sucrose intake. For the purpose of controlling the intake of sugar substitutes, people prefer to use them in specific people, such as people with diabetes, to bring them "sweet" taste enjoyment. Compared to sucrose due to a lack of chemical understanding of sweeteners, it is difficult to determine their advantages and disadvantages compared to sucrose. As for the price of sugar substitute food, it is generally accepted that it is the same as normal food prices. At least one advantage it needs to have is that it is beneficial for health, while other advantages, such as low-calorie intake and not causing dental caries, are icing on the cake. In summary, based on the effective questionnaire, we conclude that the overall advantages of sugar substitutes far outweigh the disadvantages, which greatly benefits our market promotion.

Consumer communication meeting

Record： Link

In order to introduce dietary health to the general public and our project - the heterologous synthesis of stevia glycosides. We held an online exchange meeting with some consumer groups and also invited other participating teams. The exchange meeting was mainly introduced by team leader Shiyu Long (responsible for event coordination) and team member Zhiyuan Feng (responsible for laboratory management).

This exchange meeting not only introduced the content of promoting a healthy diet and additives to consumer groups from simple to deep. Change people's impression of traditional sweeteners, as well as the public safety crisis (obesity, diabetes, tertiary and higher) that some additives can control to a certain extent. Moreover, the advantages and disadvantages of our widely used sweeteners in our speeches highlight the advantages of our new sweetener stevia glycosides advantages, including high sweetness and health benefits.

We are currently integrating the feedback gained from such meetings into the project plan to obtain theoretical support for the next step of the project. At the current stage, we are conducting research and synthesis in the biological field in the laboratory. We plan to attempt to use computers to simulate actual factory production engineering after the laboratory is completed. If certain results are achieved, we will consider implementing a complete production line for stevia glycosides, using our heterologous synthesis method for production. Of course, we have also promoted our iGEM project to the consumer community and arranged some offline experiments to give them a deeper understanding of biology, especially synthetic biology.

Expert interviews

Expert A (Link Qiao)

Record: Link

The overall global market value of stevia glycosides is around 1 billion US dollars, with a domestic range of approximately 1 billion yuan and a growth rate of approximately 10%. Currently, the price of rebM is around $300 per kilogram, which is relatively high. The price of heterologous synthesis needs to reach $100 per kilogram to have competitive value. A production capacity of 20 grams per liter is required to achieve this cost. Compared to the current artificially synthesized sweetener erythritol, which costs approximately 15000 RMB per ton, the price of steviol glycosides is relatively high. At present, stevia glycosides are not used as the main sweetener but as a supplementary sweetener. Currently, the development prospects are good, and the cost of plant extraction is relatively low. However, there are many by-products, leading to higher extraction costs. Stevia glycosides, as a natural sweetener, are safer than artificially synthesized erythritol. At present, the production cost of using heterologous synthesis is not particularly high, but if there is a lack of professional screening and transformation, there will be many by-products, which will increase the difficulty of the later extraction process. To summarize, the current market is mainly limited by selling prices. If the selling price can be reduced, there will definitely be a significant increase in market share. It will grow proportionally and linearly.

Stevia glycosides have relatively stable properties, with a heat resistance temperature of around 200-300 ℃ and relatively stable chemical properties. According to current literature, stevia glycosides have certain medical and health benefits. At present, stevioside is more widely used as a sweetener than as a health product, and its literature data is not as much as siraitin. For the products currently on the market, rebM has a better taste and is more favored by the market. However, as a high-power sweetener, steviol glycosides need to be combined with other low-power sweeteners to achieve better results. For products that currently use sugar substitutes on the market, such as Yuanqi Forest, erythritol is commonly used as a side sweetener.

In theory, heterologous synthesis can achieve low cost and high yield, and the cost can continuously change by constantly changing breeding methods. Compared to plant extraction, microbial fermentation can save land area and reduce energy consumption. The difficulty of microbial fermentation extraction is roughly similar to that of traditional plant extraction. Plant extraction mainly requires the removal of plant fibers, and in terms of microbial fermentation, yeast has a high acceptance level, while Escherichia coli needs to consider the endotoxin content. As for product purity, it is mainly influenced by the selection of enzymes, and the main substances that affect purity are the by-products brought by enzymes during the production process. Small-scale production does not require a lot of manpower, while large-scale industries need to consider more costs, such as water, electricity, and labor, as well as quality and finished product management. At the laboratory level, it also does not require a lot of manpower, but it will involve the discharge of steam and sewage. There are many requirements for the production environment in the microbial production process, which need to comply with relevant national laws and regulations.

GAINS

Through this interview, we have gained a lot of cutting-edge information about sweeteners, and experts have stated that steviol glycosides have value as both sweeteners and health products. The two are on par. If new and safe experimental methods can be obtained in the laboratory to develop the synthesis of sweeteners, it will bring huge market benefits. What we are doing is not only beneficial to the market but also to the development of synthetic biology and the development of the Earth's environmental protection industry.

Expert B (Professor Zhu)

Record: Link

GAINS

Professor Zhu first stated that our project is actually an extremely attractive one and is well worth trying. In addition, Professor Zhu stated that if we want to obtain more benefits and improve the efficiency of chemical synthesis, we need to further improve the efficiency of enzyme synthesis. Especially the synthesis efficiency of key enzymes. Improving product efficiency is one aspect, and controlling toxic byproducts is another important aspect. Some blocking strategies can be used to reduce the occurrence of side reactions.

Our experimental project focuses on glycosyltransferases in terms of protein structure. We need to explore some new regulatory sites and genes. In addition, for large-scale production, we need to design some sensors to regulate the entire production line. By designing key genes to control key metabolites, the goal of optimizing the industrial chain is achieved.

The outlook mainly focuses on economic benefits, which are competitive in the market only when they are low or equivalent to similar products, which is conducive to achieving our short-term goals. That is to say, in the fermentation tank, we need to reach a relatively high concentration to bring greater economic value. Although we are currently in the laboratory stage, we will work hard to achieve this.

Finally, Professor Zhu summarized our conversation and encouraged us to continue learning in the field of synthetic biology, learning more profound knowledge.

Exchange meetings

Meeting 1: October 5th at 2:00 PM (14:00)

Organizer: Renascence Lab

Participating Teams: HS-China, HS-Shanghai, Squirrel-Guangzhou, Squirrel-Shanghai

Record：Link

At this meeting, what is different from previous exchange meetings is that we shared our understanding of the construction of sugar substitute companies gained after communicating with experts.

At 2 p.m. Beijing time on October 5th, we had four teams participate in the iGEM-related exchange meeting organized by Teacher Yin. Our four teams introduce their members and their projects to each other. And communicate with each other to propose suggestions and learn from each other's valuable resources. Given that the projects of the four teams are in different directions and it is difficult to connect with each other, we can leverage our strengths and avoid our weaknesses to understand the progress of other participating teams. We learned a lot about depression, anxiety, and amoeba parasites during the exchange meeting. Provide new ideas for our project and expand our horizons. These contents can help us recognize and understand that there are actually more urgent problems to be solved, and we need to look at the field of synthetic biology and the world stage.

Meeting 2: October 5th at 8:00 PM (20:00)

Organizer: Shiyu Long

Participating Team: Team "Healing Assistor" from Beijing

Record： Link

At 8 p.m. Beijing time on October 5th, we had a networking event with the team Healing Assistant in Beijing. Healing Assistant mainly focuses on related research on the AIDS vaccine. Although our two teams have limited academic exchanges, we have some insights into the practical aspects of human communication. Especially in the management part of commercialization, we have had in-depth exchanges and reached a unified opinion. And after discussion and discussion, we are preparing to promote our joint event on different social media platforms. Publicize the knowledge of food additives, especially new sweetener stevioside, and the content related to AIDS (AIDS) to the public. We need to show everyone the development and changes of the new era and let the general public understand and understand relevant knowledge to cope with a completely new change.

Social Media：

In order to promote our team culture concept, product design, and iGEM-related culture, we have registered accounts on multiple platforms specifically for promoting our team culture. Our team's cultural philosophy is focus, speed, and branding. In our official account, we not only use our own native language (Chinese) for writing articles but also use the global common English for translating articles. We hope that more people can see our article, understand our philosophy, and pay attention to our products. In addition to our introduction to our team culture and products, we have also posted profiles of each of our team members on social media. Without the support of every excellent team member, we would not have become an excellent team. An excellent group not only has three of our student leaders but also several of our excellent mentors. Our group, composed of teachers and students, has brought us an excellent group that maximizes our interests, providing strong support for all aspects of our team's operations.

Education

It is very important for young children to learn about dietary knowledge, which helps them establish good dietary habits. Moreover, providing children with knowledge about food additives can help them develop the habit of choosing healthy foods when shopping. And to some extent, it can popularize knowledge related to microorganisms, allowing children to intuitively see another world and expand their horizons. Of course, while providing education, we also promoted our iGEM-related content, encouraging everyone to choose synthetic biology-related content in their future studies or work.

Education scheme：

Activity 1 -

Date：2023/08/27

Record: Link

We were delighted to host an interactive workshop at Shanghai Renaisi Biotechnology Co. Ltd., bringing together parents, students from Shanghai Foreign Language School Affiliated to Shanghai International Studies University (SFLS), and the company's leadership. The focal point of this event was to showcase our iGEM project, which entails the heterologous expression of genes related to steviol glycoside biosynthesis in Escherichia coli (E. coli). Our aim is to leverage E. coli as a production platform for steviol glycoside-like compounds, presenting a natural and healthier alternative to artificial sweeteners.

We introduced our innovative approach as follows:

Our iGEM project involves the heterologous expression of genes related to the biosynthesis of steviol glycosides in Escherichia coli (E. coli), enabling us to use E. coli to produce compounds similar to steviol glycosides. The experimental principle can be summarized as follows:

  1. Analysis of Steviol Glycoside Biosynthesis Pathway: The biosynthesis pathway of steviol glycosides has been deciphered, and relevant genes have been identified. Key glycosyltransferases can be correctly expressed and exhibit activity in E. coli.
  2. BioBrick®-Like Assembly Technique: We employ a technique similar to NEB BioBrick® Assembly to assemble multiple gene fragments.
  3. Plasmids with Compatible Origins: Plasmids with compatible origins of replication can coexist within a single cell. Plasmids sharing the same origin of replication have different resistance markers and can coexist under various antibiotic selection pressures.

We introduced the advantages of steviol glycosides to the audience:

In conclusion:

Our iGEM project aligns with the global shift towards natural and healthier alternatives to artificial sweeteners. By engineering E. coli to produce steviol glycosides, we aim to provide a sustainable and safe sugar substitute that addresses concerns associated with artificial sweeteners. We are excited to contribute to the development of healthier sweetening options for food and beverages.