Team:ECUST China/Human Practices
In this part, we will introduce you to how our human practice interacts with society, and how to continuously improve the project direction from the information obtained from enterprise visits.
First of all, we sent out questionnaires through online channels to investigate the public's understanding of phycocyanin and its application in the market. According to the survey results, we invited schoolmates to DIY phycocyanin cake activity in our school. In the meantime, we were invited to participate Hi&Fi Asia-China 2021 (Health ingredients and Food ingredients) in National Exhibition and Convention Center (Shanghai), actively communicating with Zhejiang Binmei Biotechnology Co., Ltd. Combining with the baking activity results and enterprise communication, we decided the project objective is improving heat resistance and stability of edible phycocyanin. Afterward, through business visit, consumer research and relevant policy learning, we also toke some issues up with the experimental group and the modeling group, such as food safety, processing technology, production cost and so forth. With the help of professors and experts, we continue to optimize our experimental ideas and schemes.
In HP work, we also actively convey the concept of synthetic biology to the public, expand the popularity and influence of iGEM. It is important that inform the public that the role of synthetic biology in the future development and progress of the world is immeasurable.
The work of HP is the link that connects the whole team and complements each other with each department to form an organic whole, which takes us further from our dream---Romantic blue spreads in every corner of the world.
In daily life, we find that there are more and more kinds of food containing phycocyanin in China, but their price is very high. From our data collection, we know that the cost of the existing process to produce phycocyanin about 50-90% comes from purification. We wondered if we could use green and efficient synthetic biological production to reduce its cost. After gathering many diverse ideas, we have decided to present them to Pro. Fan Jianhua，he conducts basic and applied basic research on bioactive substances of eukaryotic microalgae, cyanobacteria and other photosynthetic microorganisms.
We want to take the edible yeast Saccharomyces cerevisiae S288C as chassis organisms, and introduce the genes which have been modified to produce phycocyanin.
This discussion with the professor was an eye-opening experience, he affirmed our idea. But at the same time, he pointed out many things which had not even been considered yet. At the beginning, we just wanted to make phycocyanin in a new way to reduce the cost. He suggested that we should do more research on current market pain points and grasp its deeper problems.
Before starting our human practice to further research, we consulted professor Yang Jun , who is major in sociology. He gave us some suggestions about official requirements of iGEM, for example, how we interact with humans in the project and pay attention to personal privacy (the informed consent of collecting personal information and Opinions). Besides, he gave us some professional advice on the design of the questionnaire. It greatly promotes the implementation of Human practices. Our subsequent human practices are carried out in accordance with relevant principles.
One of our hidden consumers is the general consumer, and the other is the merchant using phycocyanin/yeast. In order to better understand the public's attitude towards the current blue food color and phycocyanin, we mainly visited consumers and businesses. Though we found that the phycocyanin in China is a new consumption hotspot, but hasn’t been widely used. It’s mainly used in the beverage, ice cream, but most businesses choose to use blueberries, anthocyanins as blue as the ornament of pigment (for example, Starbucks launched blue drinks 'ice fun brittle Frappuccino' using blueberries as the blue sources). But we do know that phycocyanin is the only natural blue food color which has been approved by the FAD and rich in nutrients.
When communicating with our consumers, we also found that Chinese consumers did not know enough about phycocyanin, and we also popularized the related functions and applications of phycocyanin to them.
In our introduction with Jia Yi bakery, we were trying to make yeast produce blue food color, which can be directly used in food bread baking. They said it was a very interesting idea and they thought it would make their food producing easier. They were very interested in our blue yeast and they thought it was a very interesting idea.
But we also found that when people heard about biosynthesis, they were very concerned about it. After briefing customers and businesses on our project "cyanobacteria", everyone expressed their doubts about our genetically modified foods. Fortunately, when everyone learned that our method of producing "cyanobacteria" is safe and can be reduced to a certain extent After the cost, most people showed a positive attitude towards our project again.
● Which way do you know to make food blue?
The results show that the public use natural food color. Phycocyanin belongs to natural blue food color which means its application market is broad and has great research significance.
● What do you know about phycocyanin extracted from Spirulina?
It’s widely known that phycocyanin has been used as a food color, but its health care function and application as a cosmetic pigment and biochemical fluorescent tracer are less known.
● What kind of phycocyanin product would you like to eat?
Analyzed by the result, our potential customers have higher expectations for the combination of phycocyanin in cakes and drinks. So we chose to produce phycocyanin with yeast, which makes it more convenient for baking and food processing.
● What do you value more when buying food? (customers)
● What aspects do you pay more attention to in the production of food ingredients?
From the customer survey results, safety ranked first, followed by price; In the processor survey, safety, production cost and market preference are factors they attach great importance to. It’s obvious that the factors customers and processors concerned a re happened to coincide. In the follow-up project design, we will keep improving to make our products close to public expectations.
● Would you like to try phycocyanin added food that belongs to similar products but has more beautiful appearance and nutritional value?（Security guaranteed）
● What do you think of the food yeast producing phycocyanin? (Natural phycocyanin is extracted from Spirulina platensis, but the cost is high, resulting in high price in the market.)
We have always attached great importance to consumers' attitude towards genetically modified food. According to the results of the questionnaire, 7% of people refuse genetically modified food, which shows that our popular science on genetically modified food needs to be expanded to eliminate people's wrong cognition of genetically modified food, and most people can accept genetically modified food with safety guarantee, In the production process of our products, we will focus on food safety and produce products that people can eat at ease.
● What do you expect from food pigment products and our project?
As a non mandatory question, we have received a total of 123 entries, and most of them have expressed their expectations for our products. What is of concern is food safety and price. For this, we will give you a satisfactory answer as the experiment is carried out and HP is launched.
In previous investigations, we learned that phycocyanin is not well understood by the public in China. In order to let participants have a close opportunity to understand phycocyanin and our project, our team held the activity ‘baking with phycocyanin’ in the community. Through showing and explaining the uses, source and efficacies to let all participants have a detailed understanding of phycocyanin. Then, they baked cake witness the cake turn blue under the spell of phycocyanin.
"I learned a lot from this activity. It was the first time I learned about phycocyanin and discovered its magic, especially in the process of cake baking. It showed me the beauty of blue. I also know how interesting thing is the phycocyanin you are making. Hope to see more of your results in the future!" Jia said.
In June, our team was invited to participate in the 23rd Natural Raw Materials and Food Ingredients China Exhibition in Shanghai, and communicated with relevant phycocyanin production enterprises in China. They are also one of the stakeholders of our project. We learned about the current phycocyanin production, the basic market situation and the bottleneck problems have been encountered in the development process. We also communicated with enterprises about our project. We have communicated with Zhejiang Binmei Biotech Co., Ltd., Henan Zhongda Hengyuan Biotech Co., Ltd., Zhuhai Yafu Xingyuan Food Industry Co., Ltd .
● Zhejiang Binmei Biotech Co., Ltd
It is indicated that phycocyanin has good color rendering effect
as a blue food color and is the only natural blue food color recognized by foreign countries at present. The
phycocyanin produced by their company is mainly sold to foreign countries, with an estimated annual sale of
10,000 tons, indicating a broad foreign market. However, phycocyanin, due to its poor stability, will slowly
fade when it exceeds 40 ℃. In the food sterilization process, the temperature should reach 100 ℃, so its
application is limited.
● Henan Zhongda Hengyuan Biotech Co., Ltd
The current application problem of phycocyanin is heat
resistance, so it is mainly used in candy, beverage and other foods.
● Zhuhai Yafu Xingyuan Food Industry Co., Ltd
Phycocyanin is expensive at present, but its
application
prospects are very broad. It is the best choice for blue food color, which is both colorful and nutritious.
But the problem is that it's not heat resistant which means the blue color will fading at high temperatures.
But he also said, "if the price of phycocyanin can be lowered, it is possible to compensate for the fading by
increasing the dose."
According to the feedback of enterprises, we know that there are two major problems in the phycocyanin market at present. One is the low stability of phycocyanin and the other is the high price. Therefore, our project solved these two problems by introducing phycocyanin related genes of heat-resistant cyanobacteria PCC6715 in the design process to improve its heat resistance.
Later, we also reported the latest situation of our project to the above enterprises and introduced the advantages of our project “phycocyanin yeast" to them. "If the results of our project can be translated, it will have a bright future in both the phycocyanin market and the food production market, especially have significance on the healthy food life of human beings," they said. All of them expressed their great interest in our project and are willing to have further exchange and cooperation in the future.
To verify that our idea is theoretically feasibleand meaningful, we consulted professor Xie Jingli, who is well versed in food engineering. And she approved us of our idea about using yeast as our host to produce phycocyain because it can be easily used to expand the production field and just using algae to expand the production field may counter with the high cost of photo-fermentation cylinder in fermentation or possible threat from the bacteria in the farm environment. Also she suggested us to use site-directed mutagenesis technology to specifically improve the heat-resistant property of phycocyanin and we should put more attention on the food safety in our project.
This set us thinking more deeply about how can we enhance phycocyanin’s heat resistant property. And we know that usually the improve of the heat resistant property accompanies with the decrease of catalyzing velocity, so we have to take more insight into the site-directed mutagenesis method. What’s more, the edibility of the phycocyanin is vital importance. Using synthetic biology to construct the metabolic pathway on the yeast, we should take care whether the source of the gene and the host are all edible and is proved by FDA or GRAS.
We have reached a long-term cooperation with Zhejiang Binmei Biotechnology Co., Ltd., which was established in 2013, is a natural extract research and development, extraction production and import and export trade-based export-oriented technology enterprises, is the domestic specialized industrialization of the production of algal cyanin-based enterprises. In August, we were invited to visit their production plant and communicated with Mr. Zhang Bing, the technical consultant. In the communication, we discussed our project and gave us relative references and long-term develop insights. We have learnt more about the wide range of phycocyanin applications, and they also have given us relevant support in terms of technology and funds.
During the visiting , Mr. Zhang Bing said, "phycocyanin is a product with great advantages in all aspects, especially in meeting people's needs for health care and it can also be used in many ways for scientific research such as the fluorescent dye. While nowadays people know little about phycocyanin and it keep a high price for long, so the promotion of phycocyanin is slowly in China... "During the communication, Mr. Zhang Bing also explained a lot about the extraction process and application value of phycocyanin to us, letting us have a deeper understanding about phycocyanin. Furthermore, he was willing to cooperate with us, helping us to promote phycocyanin on the market extensively.
At the same time, after our communication with Prof. Jingli Xie, we also hope to feed back our ideas to the professionals of the enterprise to determine the feasibility of our idea. We expressed our idea of improving heat resistance of phycocyanin to Mr. Zhang Bing, who affirmed our direction and said, "We ourselves are also exploring multiple ways to improve the stability of phycocyanin. We have already tried chemical methods to improve the heat resistance of it but the effect is poor and it is difficult to separate and purify, resulting in high cost. If we really can use biosynthetic technologies to overcome the problem of thermal stability and expressed natural blue phycocyanin heterologous, our project can be distinguished in the market. "
Finally, Mr. Zhang Bing agreed to give us their company's product——BLUE SPIRULINA as a present which is extracted from Spirulina, letting us have the taste of phycocyanin directly. And in the subsequent color-rendering experiments, we did use this phycocyanin to predict the color rendering effect at different concentrations and it provided data support for modeling .
When establishing the metabolic network of yeast, we found that some metabolites were not considered, and the flow of phycocyanin, our target product, was very tiny. Then, with the introduction of the professor, we consulted Lu Hongzhong, a professor from the school of life science and technology of Shanghai Jiaotong University. He suggested adjusting the constraints of key reactions in the original network, such as:
Besides, he also suggested that we pay attention to whether the precursor synthesis path of phycocyanin is blocked, such as protoporphyrin, protoporphyrinogen, coproporphyrinogen III, etc. We have pondered over the Lu' suggestions and improved our network. We are very pleased about the result.
During the literature reading, we found that there exists an common problem that yeast slurry was waste directly after the fermentation which was an economic loss. This made us think whether modified yeast can be applied into the fermentation process of beer to tackle with the problem of yeast slurry waste. We proceed from here and further visit school’s beer fermentation plant. In the process of the visiting of our school’s beer fermentation factory, Pro.Wang Xuedong introduced us successively of the function and order of each fermentation tank. In beer fermentation process, malt, water and hops will be introduced separately into pulverizer, gelantinization tank, saccharifying tank, filter tank, boiling pot, cyclotron precipitation, cyclotron precipitation, cooling bath and then anaerobic fermentation tank to produce beer finally.
After knowing our interest in building a plant based on beer fermentation factory, he further gave us many data based on the beer factory such as the control temperature and PH of each of the fermentation tank. Also he pointed out several problems of our yeast being used in the beer fermentation factory. Firstly, the fermentation for our yeast to produce phycocyanin calls for the aerobic environment while beer fermented in anaerobic environment. What’s more, in the last step of beer fermentation, there exists an step to filter the yeast, ensuring the taste of beer. While our phycocyanin was expressed in the cytosol, which may face it’s destination to be filtered out. The questions he pointed out guided us to further explore our hardware’s feasibility and help us get our initial hardware in shape.
We finished iGEMathon 2021 with UParis_BME.
iGEMathon concentrates on creating a novel online iGEM Human Practices & Integrated Human Practices, Ethics,
Research Journal LifeStory.
iGEMathon has three phases.Phase one is a classic involving and delving into iGEM’s Human Practuces.Local people solving local and international challenges with other people is at the elemental core of Human Practices, besides we also answering questions ranging from what we think Human Practices is to responsibility in science and scientific research.
In Phase two, we took on the task of working on a iGEMicit (creativity, innovation and teamwork) option, which means we completed a literature in the form of a fictional story——"Search the ocean"
Phase three focuses on iGEM’s peculiar Integrated Human Practices alongside answering questions ranging form "what it is” to “the process by which the purpose, design and execution of your project has you to conclude whether what you set out to do would or would not work to the end desired by you".
This task also recorded the journey of our project. We not only strengthened our team's comprehension of our Human Practices but also improved and bettered it as a result.
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IGEM_ECUST@outlook.com
ID:IGEM_ECUST
ECUST_China
East China University of Science and Technology
Shanghai,China
