Integrated Human Practice
Overview
In our project, integrated human practice plays a crucial role in helping us navigate the true demand lied under phenomenon since our ultimate goal is to shorten the time of making fruit wine and make the process economically beneficial.
To obtain a complete view of the wine market in China, we contacted some experts in the drinks and factory fields. Furthermore, we read the past policies meticulously, especially in pectin effluents. In the gene-editing techniques interviews, we learned that even though CRISPR can’t be applied to disease treatment, it is still efficient in modifying yeast’s genes. Also, the gene-modified tools used in food processing won’t cause panic among people.
CODcr index (the level of organic matter) and SS (suspended solids) index in the wastewater exceed the limit when the factory processes wine. Therefore, the use of saccharomyces cerevisiae with pectinase function can make the wine clearer and reduce pollution, to cut the cost of wastewater treatment of factories. Knowing that the public hasn’t realized the economic benefits and environmental benefits of the yeasts with pectinase, we devoted a lot of time and resources to contact all sorts of people, asking about their opinion of our project, and receiving suggestions and ideas for improvements, we develop an online questionnaire to comprehend the situation of drinks market.
Our brainstorm map is attached here：
We hope to explore the potential of our program in the fruit wine industry. Our genetic engineering experiment combines saccharomyces cerevisiae and pectinase, offering the wine industry a more affordable and sustainable solution to drinks production. To obtain constructive feedback for our project, we collected more than 1000 reports and interviewed 6 experts from the biology field and wine industry. In addition, we introduce our projects and spread our ideas to the students in the community in Shanghai.
Stakeholders
Interviews with a professional wine-maker
We are unsure if it is expensive to degrade pectin in wastewater and how important it is to reduce pectin in wine, so we asked the wine dealer Helmut Unfried some questions about the production method of wine. He replied that the importance of pectin is limited in making red wine, for pectin itself has no room for fundamental changes. Unfried said that there are more than 500 kinds of yeast globally, and each winemaker will make his wine with his special yeast. Therefore, yeast brewing is vital, and pectin is an essential part of converting into sugar.
Finally, according to his answer, the annual demand for wine yeast depends on different geographical locations and age groups. For example, the price of each bottle sold in China is between 1-10000 euros, and the customer groups that can accept these prices are also different. Investment also depends on whether it is for collection or conference.
Questionnaire
Our first questionnaire aims to survey the frequency people drink alcohol. Only 15.03% of people drink alcohol 3 times per week (the highest frequency in our questionnaire). Although nearly 50% of interviewees rarely drink alcoholic drinks, half of them who drink three times per week think drinking alcohol is beneficial. These data indicate that alcohol has a small but stable customer group.
We also investigated the relationship between interviewees’ jobs and the kinds of wine they usually drink. 42.42% of people who drink fruit wine are office workers, and the second largest group is students. One of the causes of this situation may be the tremendous pressure those people faced. Both adults and teenagers prefer beer and fruit wine to other drinks. Moreover, teenagers’ favorite drink is a fruit wine. Therefore, it is reasonable to predict that the most potential increase will be in the fruit wine market. Fruit wine will inevitably dominate the market one day.
According to the bar chart drawn by the questionnaire, fruit wine is the most prevalent in different kinds of occupations (except in entrepreneurs, who mainly drink white wine for it has higher status in Chinese drinking culture). And most kinds of fruit wine will be produced with wine yeast. It is the evidence that shows the importance of our project. If the project succeeds, manufacturers' costs will decrease, which will promote the drinks market to develop.
We also ask the question of the wine people like and the price they can accept. During 20-50yuan, it was evident that the disparity was not clear enough. But when dealing with the costs between 51-100 and above 100, we discovered that more people think fruit wine should be cheaper. So if our final yeast can help reduce the production costs of fruit wine, the fruit wine prices may be lower. And then our customers may find the prices of fruit wine more acceptable, resulting in better sales.
We further investigated the interviewees’ opinions on which gender suits fruit wine. 45.85% of people believe that fruit wine suits males, and 95.2% claim that fruit wine suits females (Multiple choice). These results can be attributed to the gentle taste of fruits wine. Contemporary women have a strong sense of self-protection. The low alcohol content of fruit wine, even if excessive consumption, will not be the same as excessive consumption of white wine resulting in stomach burn or emergency hospitalization. For contemporary women, getting drunk in a bar at night is a dangerous thing. The fact that it is difficult to get drunk on fruit wine ensures one’s safety.
We also investigated the public's opinion on which factors they will consider while drinking. 8.56% people focus on the feelings while drinking, 24.9% of interviewees care about whether it is healthy or not, 35.4% people mainly focus on the taste of the wine, only 6.61% people think about whether wine has the collecting value, and 24% people believes that different occasions decided the kind of the wine to drink. The unique and fruit flavor taste is one of the essential reasons fruit wine is so famous. According to the graph, most people consider the taste of the wine as the most crucial reason while drinking, which means our fruit wine market has a bright future.
We surveyed people about whether they know that pectin harms the environment and their opinion on whether they would still buy it if it had a negative effect. 44 people chose that they knew there was a negative impact. 51 people said they did not know that pectin had a negative impact on the environment. 125 people mentioned that they knew that pectin harmed the environment. More than 75% of interviewees realized the essence of This data shows that people have a clear review of pectin’s influence on the environment.
According to our previous research, some manufacturers add thickening agents to natural pectin, which can be harmful to both humans and the environment. From the table, we can conclude that few people are aware of the adverse effects of pectin on the human body and the environment, so it is necessary to popularize the dangers of pectin with thickeners for the general public. At the same time, the multifunctional yeast pectin that we have studied can significantly reduce the harm to the environment as it was to human beings.
We investigated people’s acceptance of gene-editing becoming mainstream of processing food (in final food). More people chose to reject the gene-edited food. We can see from the data that if we stressed the gene-editing techniques applied in our last project, we might get re- refusal from the general public. The acceptance of gene-edited food into mainstream food is not ideal. We need to reconsider how to balance the promotion of our product and the authenticity of academic technology.
We surveyed people's opinions on "whether it is acceptable to use gene-editing technology to process tools for food production （Not included in the final product）." Among them, 149 people chose "acceptable," and 71 people chose "unacceptable." The total number of acceptable people was more than twice as many as the number of unacceptable gene-editing tools, indicating that people are more accepting of gene editing tools. Most of them can accept that the final product does not contain gene-editing technology. This reminds us wine should be labeled that it is free of gene-edited products.
Reasons, why people drink fruit wine should be the main factor we are concerned about. Our investigation found that 36.15% of people drink fruit wine because of its great taste, and 29.67% drink fruit wine because of low alcohol. And our products have the functions of improving the taste and reducing the sourness fruit wine contains. So it will fit the expectations people have on fruit wine.
Experts’ Opinions on our Techniques
1. Interviews
To further investigate the characteristics(both advantages and disadvantages) and racial problems of CRISPR in synthetic biology and improve our product, we have two rounds of video interviews with Professors in both gene-editing areas and enzyme engineering.
First stage
Dr. Yibei Xiao is a professor at China Pharmaceutical University. He chose to apply for CRISPR because he thought it was the direction of the future. So he started to study CRISPR bacterial antivirals, and he switched from studying human antivirals to bacterial antivirals. He mainly studies CRISPR-CAS and has published four papers on Science, Nature, and cells.
Before the project starts, we are concerned about whether the CRISPR is safe enough to use in the food procession. And he told us the use of CRISPR in the laboratory is relatively perfect, low cost, and easy to operate. So it is relatively easy and safe to apply the technology to yeast targeting.
He also mentioned two current existing problems of gene-editing technology. One primary concern is the off-target effect, were editing a specific gene may accidentally edit other genes and cause unintended side effects: 1% of cells edited incorrectly may produce cancer. However, we are not applying CRISPR in changing human gene sequences. It is pretty safe to use CRISPR in wine production.
Ethical issues also exist, especially the public's fear of alien technology. For example, American scientists put the carotenoid gene into the rice gene in the golden rice incident to make rice with high carotenoid content. When Chinese children consumed the rice, it caused panic because there was no long-term scientific proof, and it might have side effects after many years. We get to know that directly applying gene-editing techniques to humans is too fast for now, even though there is no harm. We need to reduce public prejudice against bio-editing technology. Also, it is our responsibility to communicate to the general public the safety of using gene-editing technology as a tool to mitigate any unnecessary hassles that may arise when advertising the product in the future.
Second stage
Dr. Xingzhou Chen is a professor at Wuhan Institute of Technology. He mainly researches enzyme engineering. Two years ago, he started studying the wine-producing industry and was an "excellent instructor" in Hubei Students Wine Body Design Competition.
He pointed out the problems in the initial wine-making process. Wastewater that concludes pectin will pollute the environment, features in high COD and BOD index. It can also block the winery's blow-off line and create an extra clearing expense.
"It is technically feasible, and the CRISPR technology you use is relatively mature. The yeast fermentation technology is also very mature now." When we asked him about his opinion on our experiment, he answered.
He added, "When CRISPR causes DNA double-strand broken strand recombination, it is not completely random. It is biased. If it is completely random, then we cannot use it, and the efficiency is extremely low. Since CRISPR has been used until now, it still has its advantages, so there will be no large-scale disorder."
We learned two points they have to be careful of when in the laboratory and applying our product.
1. While commerce, chateau must be informed to remove pectinase from the final product. This process can relieve the public's panic since gene-editing is only used as a tool and will not be included in the final product.
2. The yeast is used under aerobic conditions in the lab, but alcohol fermentation is un-der anaerobic. Therefore, experiments must also be carried out in anaerobic conditions in the laboratory to make our results more practical. Later, we told the experiment group to use these aerobic and anaerobic conditions in the laboratory.
We mentioned that DNA double-strand break repair mediated by CRISPR-Cas9 is very likely to cause the loss of large DNA fragments at the distal end of the cutting site, which can be located be- yond 20kb. Moreover, this negative effect is highly random, and the edited genotype of each sample is different.
This means that using CRISPR technology to edit genes is prone to more severe structural variations such as gene translocation.
In turn, the professor tells us we must clear regulatory mechanisms. Initially, we must know how many genes are in a piece of the genome and what relationship is between these genes to complete the subsequent experiments.
This information gave us both instructions and restrictions for the experiment. We mainly need to focus on our projects' environmentally friendly aspects during the production process and advocacy.
Third Stage
We interviewed an expert in fruit wine manufacturing technique, Xiaoyu Han. He is less well-known but has done profound research in zymurgy.
At the beginning of the interview, he informed us of two documents that will be helpful to us: Guidelines for the development of the 14th Five-Year Plan of China's liquor industry and GB15037-2006 (national standard). The former points out that the fruit wine market is already expanding at approximately 3% a year and has a bright future since teenagers will become primary consumers in the future. The latter regulates dry extract content (including tanning, pectin) must exceed 18g/L in red wine. This corrects our misunderstanding that the final product must be pectin-free.
We asked his opinion on the practical significance of our product. He mentioned that 14 years ago, gene-edited yeast already existed in the market, but due to strict control of gene-edited products in Europe, there wasn't a large-scale application.
More factors will influence the practical significance of our product other than the government's restriction. First of all, gene drift will influence the gene's stability. For example, there was an experiment on gene-editing beer yeast, making it produce aroma substance without using hop. Although the experiment had a good reflection in the academic field, it wasn't successful in the industrial circle since it has low stability and high cost. The quality and quantity of aroma substance it produces are insufficient too. Secondly, gene escape may also disturb the chateau's ecological environment. This inspired us that we have to consider the efficiency of the pectin decomposition of our product. Moreover, we will better understand yeast's physiology and fermentation process in future research and find other meaningful research directions. In conclusion, our experiment is only a starting point. We need more effort to expand it and make it practical.
He further explained that although pectinase's cost is high for the chateau, not all wines need pectinase while producing. For example, the grape has thin pulp cells and doesn't need pectinase while producing wine. This explains why red wine chateau owner Unfried thought pectin is of little importance while making wine. However, other fruits with thick pulp cells such as peach, kiwi, blueberry, and mulberry need pectinase while producing fruit wine. This information indicates our potential customers and abandons corporation with red wine chateau.
In previous research, our team had an idea of adding phage's gene into yeast to have a better antibacterial property. He mentioned that deterioration mainly results from acetic acid produced by acetic bacteria, and some current saccharomyces cerevisiae already has an antibacterial property. Therefore, he suggested we aim at the antioxidant of our product. The current oxidation resistance method is adding tannin. If our multifunctional yeast has the antioxidant function, production costs could be cut too. Therefore, we can do a further experiment on this topic.
Fourth Stage
After the interview with Mr. Xiuxuan (Atlas) Zheng, senior consultant at Oriental Securities, we sent him our business plan to ask for advice. Mr. Zheng said our business plan should be more concise, extracting the core concepts and using chart analysis. The detailed content can be presented on the website. Then, we need to condense our point into a single sentence, which can be bullet points. Using data tabulation, our views and ideas will be more intuitively reflected.
We need to let the public know where the greatest value of our products lies, and at the same time, let our target customers know what the advantages of our products are compared with other products.
Field trips
On-site Investigation to Manas Company
On 11th August, we visited the Manas wine cellar. The winery we interviewed is mainly used for sales, and the staff only has a superficial understanding of overall winemaking technology.
The employee thought that the most complicated and costly part of the winemaking process was the fermentation stage and that he would choose to buy new yeasts that could ferment wine and break down pectin if they were developed.
There is also competition in the way of winemaking. Machine winemaking is cheaper but the taste is worse and is not good for health; traditional oak barrel winemaking is more expensive but has a mellow taste and is conducive to people's health. For Manas, the most important way to improve the reputation of Manas is to enhance the quality of the wine.
For oak barrel winemaking, it usually requires a longer time for fermentation. If the quality of wine and the shortening time of making wine are compatible, it is not hard to imagine that the gene-editing yeast will be prevalent in the current market. Also, our yeast improves both the fermentation stages in machine winemaking and oak barrel winemaking. As a result, we will face a larger market than we previously assumed.
Key Conclusions
After completing the human practice activities, we obtained several key conclusions.
1) Fruit wine has a great market potential in China;
2) For fruit wines pricing over 100 RMB, most people consider that the price should have been lowered;
3) It is quite safe to apply CRISPR-Cas system to modify yeast, whereas, experiment safety issues should be noticed;
4) Since most people do not support the application of GMO techniques in final food, we should be cautious in promoting our gene-edited yeast to the general public.
5) A business plan should be concise and eye-catching, stressing on the major competitive edges of our products.
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