Team:UCAS-China/Human Practices
Human Practices accompanied us through the year, as DeCaffi is such a unique project with brand new concepts of managing caffeine intake and difficulties of modifying and providing food enzymes directly to end users, HP activities help us to make sure we are on the right track. Through HP activities, we determined the main concept of our project, decided on details of the design of the project to reach for the best and had a brave envision for the future.
Beginning
During brainstorming, DeCaffi was a competitive candidate for 2021 iGEM, its value and meaning were revealed through adequate investigation.
Questionnaire survey
Aimed to obtain a comprehensive understanding of the intake of caffeinated beverage and public perception of genetically modified technology and enzyme products in China, we designed and placed a questionnaire containing 17 questions about caffeine, enzyme preparation and genetically modified products. Fortunately, we received 410 effective questionnaires from 10th, June to 3rd, July. The following is a brief analysis of some critical parts in our results.
I. Sample Distribution
Fig.1 Age
We can find that nearly 80% of responses were comprised of those aged 10-29 and 40-49(Fig. 1), who are exactly the target groups of our project DeCaffi, as the main consumers of caffeinated beverages that we envision. The samples aged below 10 or beyond 60 were excluded for the rest of the analysis due to the lack of quantities.
Fig.2 Education
Additionally, the data shows that a majority has received high school education and higher (Fig. 2), which may have an impact on the results of the awareness survey about GMOs and enzymes.
II. Caffeine intake and cognitive profile
Fig. 3 Reasons for consumption of caffeinated beverages by people with different drinking frequency
Through the data, 70% of the respondents have a habit of drinking typical caffeinated beverage like coffee, tea and milk tea every week, which means our project has a wide range of potential target people.
Fig. 4 Percentage of caffeine-sensitive people
The figure above reflects that caffeine-sensitive people account for 18% of our respondents (Fig. 4). Combined with the phenomenon that the majority drink caffeinated beverages in order to enjoy the flavor rather than keep awake with caffeine (Fig. 3), it’s necessary to find a solution for those who are kept away from their favorite caffeinated beverages due to caffeine sensitivity. And DeCaffi gives a possible version. Meanwhile, the importance people attached to beverage flavor reminds us to pay attention to the influence our products have on the flavor.
Fig. 5 Knowledge of caffeine content in beverages
Although almost everyone cannot avoid caffeine intake, over half of the respondents have no clear understanding of the potential harm caused by caffeine to the human body (Fig.5). From this perspective, we’d like to create a portable cup to let users know the caffeine content in their beverages (more details on hardware page). Furthermore, we attempt to shoulder the responsibility for public health awareness promotion, which is also a core concept of DeCaffi.
Fig.6 Awareness of caffeine management
In addition, we conducted an intention survey on WeChat applets to get a general idea of what services people expect in our applets. More details in this aspect can be found on our software page.
III. Public awareness and attitude towards GM technology and enzyme products
Fig.7 The popularity of GM technology and enzyme products
Public attitude towards GM technology and enzyme products is an enduring topic. It can be clearly seen from the figure that the concepts of GM technology and enzyme products have been popularized among the public, while their specific application scenarios still need further popularization (Fig. 7). As for public attitude, enzyme products have a higher acceptance level than GM technology (Fig. 8). Since our project use genetically modified bacterium to produce enzymes we need, we tested people’s acceptance level of it likewise and it is pleased to find that over 90% of people accept it.
Fig.8 Public attitude towards GM technology and enzyme products
Theacrine Investigation
In order to degrade caffeine in a portable and ecofriendly way, we chose a natural metabolic pathway found in a Chinese plant Kucha that transfer caffeine into theacrine. To ensure the availability and safety of introducing theacrine into coffee, we communicated with Mr. Hu Dan, who participated in the identification and characterization of CkTcS. He believes that using CkTcS is a new direction for production of decaffeinated drinks. In the communication, he commented that our idea was creative and had great potential, while pointing out that one key point of our project is making sure that Cdh and CkTcS can efficiently working together.
We have conducted a sufficient literature reading to understand the function of theacrine on human body. According to toxicological studies on rats and mice, the toxicity grade of theacrine was determined to be non-toxic(Xie, 2009). Other experiments on rats and mice implied that theacrine have sedative-hypnotic property(Xie, 2009), anti-inflammatory(Xie, 2009) and antidepressant effect(Xie, 2009), and may be able to reduce liver steatosis.
In The Process
When we tried to introduce our project to our fellow students, two questions were often asked: “why would I drink coffee if it cannot keep me awake” and “why would I drink coffee if I cannot tolerate it”.
Why? Why should people choose decaffeinated coffee? Why should people choose our method? What can coffee bring to us when it no longer keeps us awake? Our Human Practices focused on these questions.
Enjoy coffee with a good night’s sleep
By attending the Tianjin Coffee Mart and visiting several cafes, we learned that there is a little but constant demand for decaffeinated coffee from those who love enjoying coffee but do not want to stay up late. A worker from Starbucks Reserve told us that more and more people regard coffee as a drink instead of a refreshing potion. They went to a coffee shop with their friends, tried their first cup of coffee, and then fell in love with it. Mr. Tim from Manner Coffee expressed the uncomfortableness of longing but cannot drink coffee at night.
These conversations made us more determined on expanding the probability for people to try their first cup of coffee without worrying about their tolerance of caffeine, and expanding the time in a day when people can enjoy coffee.
Decaffeinate without hurting the flavor
A worker from Starbucks Reserve told us that the cost of a traditional decaffeinate method-Swiss Water Process is low, but it may hurt the flavor of coffee. Coffee maker from the Metal Hands told us that the process is usually not used on high level coffee beans. At Berry Beans, we tried the Colombian decaffeinated Coffee，who had a sour bitter taste, and the smell was less attractive. Coffee roaster Mr. Ma had the same opinion, he believed that a decaffeinate method that does not hurt the flavor would be favored by coffee lovers.
By making coffee flow through immobilized enzymes Cdh and CkTcS, we can change caffeine to theacrine, we can directedly degrade caffeine without hurting flavor compounds, as a result we can preserve the flavor of coffee as much as possible. However, existed in Kucha, theacrine may produce its own taste. Worker from Starbucks Reserve and coffee maker from Metal Hands both agree that trying to develop a special coffee is a good choice.
Manage caffeine intake
Coffee industry in China is developing rapidly. Covered in the third coffee wave, thousands of coffee shops appeared in China, where every links from planting to picking, processing to transportation, baking and brewing are strictly monitored to ensure high quality. We were not sure if DeCaffi can perfectly fit into the thriving coffee industry, while a conversation with manager of Half Half Café opened our mind. “Nowadays, what is on the shelves should not be products, let alone technologies, but culture.” Being inspired, we set out to try expand DeCaffi from a method to a concept: managing our caffeine intake and every part of our health as we manage our exercise, water intake and sleep.
We sent out questionnaire to understand people’s awareness of caffeine and their sensitivity to caffeine on average.
While caffeine containing beverages have become the “water of life” of modern people, we believe DeCaffi will be a perfect solution for people to learn about caffeine and manage their caffeine intake. Guided by this concept, we successfully invented a smart cup and a smart app.
Compatible in various caffeine containing beverages
To reach the goal of managing caffeine intake, we need to expand the usage of our project from coffee to tea, milk tea and other caffeine containing beverages. Therefore, we abandoned the previous accompany cup design and chose to construct a set of suites that can be used in various kinds of cups. We modularized the filter element, spoon and detection devices. Now, we are trying to miniaturize and integrate the electronic components in the suites to expand its application scenarios. (more detail see hardware)
Safe
Cooperated with BIT-China, we thoroughly investigated current Chinese law and regulations on food enzymes which encompassed us to design the project to meet the standards.
| Law or Regulation | Requirements | Design |
| National Food Safety Standard | The raw materials do not pollute the final food residues. | A filter that immobilize the enzymes. |
| Use qualified microorganisms for production. | Use safe and well-studied E.coli as chassis. | |
| Ensure stability of the method and conditions of strain preservation. | Ensure proper operation of bacteria in the laboratory. | |
| The detection of pollutants, microorganisms and antibacterial activity reach the standard. | Future experiment plan. |
We also consulted Mr. Tao Yong, investigator in the Institute of Microbiology, Chinese Academy of Science, about his opinion on current law and policies on food enzymes and genetically modified food. He pointed out that an imbalance exists between scientific research and laws and regulations on GM technology. But it takes time to develop a strict and comprehensive policy and relative departments is taking action, thus we can be positive to see scientific research finally pay off.
The Future
When the Giant Jamboree is approaching, we reflected the whole project to see what we missed and what we can do in the future.
The scholar
We invited Mr. Tao Yong to watch our brief rehearsal of the final presentation and gave us some advice. Mr. Tao is experienced in microbial fermentation of enzymes and pointed out many problems that we can improve in the future.
Key Points:
- Try to achieve the recycling of the coenzymes and cofactors to lower the cost.
- As Cdh and CkTcS comes from eukaryotic plants, constructing the gene circuit in S. pombe may lead to a better result; the expression of enzymes is an art that need multiple attempts of multiple strategies.
- To meet the need of most people and factorization the project is another way to lower the cost.
The company
Established in 1996, SUNSON has grown from the first local company to produce fermentation cellulase to a full functioning enzyme company. A conversation with SUNSON brought us closer to the market of food enzymes.
A technician from SUNSON thought highly of our idea, and pointed out the difficulty and time consuming of modify the enzymes to fit in the temperature and pH of coffee. As delicate molecules that only work in a certain condition, perhaps it is not time for enzymes to become tools and means in daily life, but we believe we are on the right path. There are tens of amylase that can be used at different conditions, as it is needed in many kinds of industries, the cost of finding and transforming new enzymes has been diluted by factory production. It is possible for us to construct different Cdh and CkTcS that can be used at different daily scenarios. What we need to do in the future is to find a balance between meet the needs of most people and meet the need of individual.
References
Gao, M., Zheng, J., Zheng, C., Huang, Z., & Huang, Q. (2020). Theacrine alleviates chronic inflammation by enhancing TGF-β-mediated shifts via TGF-β/SMAD pathway in Freund's incomplete adjuvant-induced rats. Biochemical and Biophysical Research Communications, 522(3), 743-748. doi:10.1016/j.bbrc.2019.11.126
Jun, X. (2014). Effect of theacrine on lipid metabolism mediated by SIRT3 / AMPK signaling pathway and its mechanism.
QIN Dandan, W. Q., Li Hongjian, FANG Kaixing, JIANG Xiaohui, PAN Chendong, WANG Qing, LI Bo, WU Hualing. (2021). progress in research on Camellia kucha (Chang et Wang) Chang and Its Special Constituent Theacrine. Food Science, 42(13), 242-259.
WU Minzhi, H. Y., CAO Yating, LI Laidong, ZHOU Heliang, ZHU Rui, LIAO Yanfeng, LI Yuanbo. (2009). experimental study of theacrine on antidepressant effects. Chin Pharmacol Bull, 09, 1160-1163.
XU J K, K. H., ZHAO L. (2007). Theacrine, a special purine alkaloid with sedative and hypnotic properties from Cammelia assamica var. Journal of Asian Natural Products Research, 9(6-8), 665-672. doi:10.1080/10286020601103155
