For the first time we thought of flavonoids, a group of organic compounds contained in almost every vegetable, such as Chinese cabbage, orange, and haw, we found that it was beneficial to human bodies in a collection of aspects. Anti-aging effect, for instance, is one of the most significant benefits of flavone through the function of antioxidation. Besides, flavone is anti-inflammatory, as well as antibiosis, and anticancer, which is of great help to improve human health.
Nevertheless, human bodies themselves cannot degrade flavonoids; rather, a typical type of enzymes presenting in intestinal flora plays an intrinsic role in flavonoids absorption. The thing is distributions and population of intestinal flora is disparate in different human bodies with respect to their dietary habits between zero and five. Hence, some people develop enough enzymes in intestinal flora for flavonoids digestion and absorption, whereas others do not, which leads to a problem that the latter are not able to degrade and absorb this substance well. Fortunately, until 2020, NATURE COMMUNICATION published a paper reporting the key enzyme FLR that catalyzed the first step of flavonoid degradation, which can help degrade a variety of flavonoid compounds. Therefore, our purpose is to inject the gene fragment of FLR enzyme into e-coli that can better express FLR gene, improve degradation of flavonoids, further produce DAT and stimulate the immune system of human body, and reduce clinical treatment costs at the same time.
First and foremost, in order to standardize our cognition to public understanding of flavone and its function, we launched a field survey in Lvdi Square, designed an online questionnaire, and sent it to the public.
It can be shown from the response in field survey and online questionnaire that a large portion of the public have a poor understanding of flavone as well as its benefits except those who are major in Biology and Nutrition. As a whole, consumers focus more on validity, efficiency, and safety of our new products compared with their price, and over a quarter of them look forward to the great prospect of our products. However, we should also notice opponents beneath the surface covered with support. Some, for instance, cast doubt on potential risks——including gynecological diseases caused by estrogen overdose——of the product. A few of the population who have limited comprehension of what flavone and its functions are showed indifference to flavonoids commercialization. Thus, it is essential to publicize its function to the public and offer solid and forceful replies to the skeptics to raise public awareness, since questions and widespread ignorance are two major obstacles that restrict the development of our products.
To have a better understanding of our project, we interviewed a foodstuff safety expert and a Bioengineering expert for more information. The former informed that due to the structure and characteristic of flavonoids, cooking at high temperature will lead to rapid oxidation and the loss of biological activity of flavonoids. What is more, there is little evidence of excessive intake of flavonoids. The latter three factors which determines whether injecting FLR enzyme into e-coli can convert flavonoids into DAT and activate human immune systems, which makes human bodies able to absorb flavonoids in an easier and more stable way. Moreover, he provided support and extra information for our further experiment and research.
Associated with the function of flavonoids and the improvement of flavonoids absorption through FLR enzyme injection, we choose people who trouble degrading and absorbing, especially flavonoids, people who looking for anti-aged products to reduce the rate they age, and children and adults who just like yoghurt as our target group.
The principle of our project is that we inject the gene fragment of FLR enzyme into pET28a, an e-coli that can express a certain enzyme efficiently to form an engineering bacterium containing FLR enzyme. This strain can better express the FLR gene, improve the degradation of flavonoids, further produce DAT, and stimulate the immune system of human bodies to achieve anti-inflammatory, antibacterial, anti-cancer and other purposes, at the same time to help reduce clinical treatment costs.
As we use flavonoid degrading enzyme, its degradation effect is very considerable. Our main goal is to replace those gastrointestinal bacteria with this flavonoid degrading enzyme. Although those bacteria have co evolved with humans for many years, we chose FLR enzyme to degrade more flavonoids in order to play the role of flavonoids perfectly. To achieve the purpose of producing dat beneficial to human body after flavonoids degradation. Our data showed that the average residual flavonoids of about 0.198mg/L were obtained after the degradation of different varieties of flavonoids with an original content of about 10mg/L by our FLR enzyme. This shows that our FLR enzyme plays a significant role in the degradation of flavonoids.
Our products will be produced as powder, which can be stirred with yoghurt, or other liquid drinks. Besides, every 200 ml of them will be packed with plastic, which is cheap and is mass produced. However, the price of the product itself, among 8-12 RMB, is relatively higher than the market price of common yoghourt, which is 5-12 RMB.
Considered of the competitors in the market, operations that sell yoghurt and probiotic powders and probiotic drinks are firms that we ought to take into account. If we delve into their pros and cons, public understanding and low price are two major aspects of their advantages. Consequently, it is clear that we need to work out the solution to producing our products as powders with relatively a higher quality in the same quantity——that is, a higher validity to absorb flavonoids. Furthermore, an efficient approach to advertise and publicize is also important. In addition to public speech, creating a website, and updating official accounts, attractive posters and necessary science popularization are required to make new customers enter the market. In order to achieve the intention of cooperation, our project team seek high-profile and influential bloggers to promote our products, similar to The Little Red Book and INSTAGRAM, so that the public can know about our products through articles or shares from bloggers, and then they will buy or invest. Moreover, through short video platforms like Tiktok, our project team can push our product advertisements to our target groups or let influential video producers integrate our products into the videos, so that the public can better accept and get interested in our products. To keep old customers in the market, our project team are supposed to offer discounts and membership, similar as Freshippo and Sam's, for the purpose of not only making our product the first priority when purchase for flavonoids degradation and absorption, but also enjoying their time with family and friends when shopping. Simultaneously, we ought to always be sensitive to changes of public demands in the market, which is of great help for us to follow the trend and keep increasing our profits.
Product Development Plan
We plan to try more different flavonoids and repeat the experiment multiple times to ensure that the error is reduced. To go further, we are looking forward to test how human body would consume with and without the pET28a-flr. What's more, if the further experiment succeeds, we will probably try to add the pET28a-flr into yogurt. Yogurt contains a large amount of lactic acid bacteria, which can promote gastrointestinal peristalsis and food digestion. Yogurt helps on digesting food and pET28a-flr also helps on digesting flavonoids. By adding it, we will probably see a win-win situation which is actually great for human health improvement.
Safety Concerns and Other Possible Challenges
The development of enginerred probiotic is actually strictly administrated by the government and other supervision departments. The safety, reliability, toxicity, and storage conditions are all worth considering. Future experiments should be rigorously designed and conducted to ensure these aspects of future development of our probitoc and related product. In addition, as mentioned in our previous point, the mechanism of our potential probiotic should be further explored. For different groups of people, the probiotic may take effect at different conditions. Our future research and development should also focus on this aspect.