Integrated Human Practice
After the outbreak of the COVID-19 pandemic, the demand for alcoholic products such as medical alcohol, alcohol disinfection has increased rapidly. Only in 2020, the output of fermented alcohol in China was 9.24 million kiloliters, with a year-on-year increase of 33.6%. In the meantime, the export volume of fermented alcohol in China also increased in 2020. In that year, the export volume of fermented alcohol was 624.484 million liters, while the import volume was 182.146 million liters.
The above data indicate that faced with COVID-19, alcohol is urgently needed in China, either to satisfy the domestic demand or to enter the global market. Thereafter, we plan to focus on alcohol production processes in the 2021 iGEM Competition. The primary goal of our project is to obtain a yeast functional in making wheat b-starch efficiently ferment and produce alcohol economically.
After gene editing, our yeast is designed to secrete three enzymes. They can accelerate the decomposition of wheat B starch into monosaccharides (glucose), and the yeast can further ferment glucose into ethanol. In the past, the three enzymes are required to be added, but now they can be secreted by the yeast naturally, which can lower the cost and shorten the time needed to produce alcohol. In addition, the output will be increased due to the wheat b-starch is taken into production. Therefore, as the supply increases, the market price of fermented alcohol will decrease.
To move forward with our project efficiently, and to integrate as many as possible stakeholders’ opinions in our project, we conducted a series of human practice activities. As a result, these activities enlightened our future direction, and further confirmed our confidence & determination to continue our project,
Our human activities are divided into two parts: Expert interviews, questionnaire surveys.
Expert Opinions
Interview with Dr. Xin Wang, Ph.D., Chinese Academy of Sciences, Professor at Henan University of Technology
Dr. Wang has obtained his Ph.D. degree from the Key Laboratory of Synthetic Biology at the Chinese Academy of Sciences. He has abundant research experience of yeast, thus, we interviewed him about the feasibility of our yeast project.
Importance of Yeast in Alcohol Production
Dr. Wang clearly explained the role of yeast plays in the brewing process and modern alcohol production. Based on his explanations, we further understood the general process that yeast mutates monosaccharides such as glucose and maltose into alcohol through a physiological metabolic pathway, which is generally referred to as the glycolysis pathway, the process of converting glucose into pyruvate in any microorganism (cell). More importantly, we confirmed that yeast has been an irreparable role in modern alcohol production because yeast fermentation produces alcohol with high purity and fewer impurities.
Discussion about our Gene-edited Yeast and Possible Impact
After a general introduction to our project, we came to seek Dr. Wang’s opinions on our gene-edited yeast. Dr. Wang thought that compared with current fermentation techniques, our gene-edited yeast may bring about extra benefits. He mainly held that currently corn starch has been used as raw material to produce alcohol, whose price has been increasing all these years and reached 3000 RMB per ton. In contrast, wheat is only less than 2500 RMB per ton, Therefore, he thought that if our yeast makes wheat starch ferment efficiently, we may dramatically lower the alcohol production costs.
Dr. Wang also mentioned the impact of our project on environmental protection. He thought that currently wheat B starch is not used efficiently used. Neither feed making nor direct blowdown causes problems, e.g. livestock health problems or environmental pollution. Thus, he thought that it is also an impressive idea to make wheat B starch as raw material to produce alcohol from the perspective of environmental protection.
Technical Advantages of our Yeast
Lastly, Dr. Wang also helped us with analyzing the competitive edges of our gene-edited yeast.
According to Dr. Wang’s analysis, we further collated the general working principles of our yeast, e.g. how our yeast interacts with the alcohol production process, why could our yeast save production costs, etc.
In the process of making alcohol, B starch needs to be converted into a monosaccharide. In the process of transformation, enzymes need to be added to produce monosaccharides, but the more enzymes added, the higher the cost. Therefore, if yeast can secrete enzymes by itself, it can save costs. Our iGEM engineering strain is an engineering strain that can autocrine and degrade the sugar in wheat starch, and change monosaccharide to yeast.
Three enzymes are needed in the process of turning starch into monosaccharides, which should be added in the brewing process, but it also costs money. So edit the gene of yeast and let it autocrine, so that it can turn B starch into sugar and then produce alcohol.
The functions of these three enzymes are used to turn starch into monosaccharides. They cooperate and have their division of labor. In type B starch, there are two types of sugar, one is starch and the other is pentosan. Starch is easy to turn into glucose and enzymes are cheap. But in this way, pentosan will be wasted, so to find a way to turn pentosan into a monosaccharide, we need to add three enzymes.
One is xylanase. Its function is to randomly cut long-chain Pentosans (connected into polymers) at random, so glycosidic bonds change from hundreds of polymers to three to five forms, so it can greatly reduce the degree of polymerization. The second enzyme, beta-xylosidase, is used to cut the polymers into about three to five monosaccharides. The last one is acetyl xylan esterase, which is the branching enzyme. Because there are some side chains (granules) in the whole pentosan, these Pentosans are not pure sugar molecules, and there are some acetyl groups. Therefore, these will affect the components of monosaccharides, so acetyl xylan esterase is used to remove them.
Therefore, the three enzymes have their division of labor, and the pentosan in starch is changed into a monosaccharide. Yeast can only use monosaccharides, not a sugar-like polysaccharide, so it's like cooking for yeast.
From an interview with Dr. Wang, we have developed a better understanding of yeast in alcohol production, and how we can lower the production costs of alcohol in an environmentally friendly way. All in all, we have obtained further confidence in moving forward with our project.
Interview with Mr. Andong Zhang, Senior Manager, COSCO SHIPPING LEASING CO., LTD
Since Mr. Zhang has rich experience in industry consulting, he may give us some insights into how the yeast-related industry work.
Mr. Zhang mainly reminded us of the safety concerns about yeast, he emphasized that we must have our yeast certified by the China Food and Drug Regulatory Commission in our future business development. Also, he confirmed that our techniques will make a difference to the alcohol manufacturing industry. In the end, even though Mr. Zhang thought that many people have a bias towards GMO products, the current market is still bright.
From interviews to the business expert, now we can establish a feasible path for business decisions and the safe handling of edited yeast. The yeast has a wide market to sell to, we must continue to improve and execute experiments and carefully make every business decision.
The interview with Mr. Zhang mainly reminded us that we should pay attention to the safety issues of our project and we needed to adhere to relevant regulations and laws in future business expansion.
Interview with Dr. Canhui Liu, Ph.D. Candidate, University of Cambridge
Since Dr. Wang helped us analyze the environmental benefit that our project may bring about to society, we were much more inspired. However, are there any environmental concerns we should pay attention to? With this question in mind, we interviewed Dr. Canhui Liu, a Ph.D. candidate at the University of Cambridge, whose research expertise includes environmental policies and sociology, This time, Dr. Liu gave us different while well-judged comments about the environmental concerns that our project may bring about.
Impact of Experiment on Environment
Wheat B starch itself will not affect plant production type, but experts mentioned that in our experiment, a lot of yeast was added. In the process of yeast decomposing wheat B starch into alcohol, will some harmful gases, wastes, and compounds to the environment be produced? Because the composting treatment of wheat B starch does not do much harm to the environment and plants, we should try our best to keep our experiment green and harmless. The catalysts and yeast we add should ensure that they do not have too much toxicity and harm to the soil or water body. For example, the sulfide and nitride produced in the experiment should be reduced as much as possible.
Impact of wheat B starch on the environment.
Dr. Liu mentioned that we should strive to achieve a waste-free city - promote the formation of a green development mode, continue to promote the source reduction and resource utilization of solid waste, minimize the landfill volume and minimize the environmental impact of solid waste. Wheat B starch belongs to industrial solid waste and rural solid waste in waste-free cities. Therefore, our topic is to reuse useless wheat B starch and turn waste into treasure, which is conducive to the realization of waste-free cities. Later, experts mentioned that if wheat B starch is not used, the treatment methods of wheat B starch are generally shunting treatment, feeding livestock, and composting. Because the nutrition of wheat B starch is low, if fed to livestock, it is not conducive to livestock digestion, which will make livestock indigestion, and livestock will also produce a lot of greenhouse gases when excreting, thus affecting global warming. Composting treatment is an optional treatment method, but the efficiency of composting treatment is too low and the cost is too high, so it will affect the economic benefit and reduce the economic benefit. Experts stressed that we should try our best to achieve carbon neutral and waste-free cities, which is very important and our goal.
What is the impact of yeast after gene editing on the environment?
After gene editing of yeast, it will have a great impact on the water body. Firstly, at the industrial level, some waste liquid may be produced in the process of gene editing and fermentation. If this waste liquid is discharged into the water body, it will certainly cause eutrophication of the water body, which will cause great pollution to the environment, Therefore, we need to treat the sewage containing molecular proteins produced in the process of gene editing yeast and decomposition into alcohol. The second aspect is that some of the remaining materials or articles produced must contain some liquid during composting. It is necessary to ensure that the liquid concentration is not too high because the liquid will further produce fermentation under natural light, which will have an impact on plants.
What should we pay attention to in the experiment?
We should treat the wastewater and some polluting substances produced by the experiment.
As an expert in environmental policies, Dr. Liu reminded us that safety should be placed in the first place throughout every step of our experiments and project. We must treat our project from an objective perspective and cautiously proceed with it.
Interview with Mr. Zhenfeng Su, Investment Manager, GoHigh Capital
The last expert we interviewed is Mr. Su, who has quite a lot of experience in business planning and investment. He gave us great frameworks about how to design our future implementation plan.
Commercial interview summary:
In summary, Mr. Su explained how to write the business plan, including the structure of the business plan and what is supposed to involve in each part. Secondly, he answered our question of what our target consumers are, he mentioned that our target consumers are mainly alcohol manufacturers because the product our company sells is genetically edited yeast, retail consumers do not have the requirement to purchase such yeast. Then he gave us suggestions about what marketing strategies should be supposed to undertake when our target customers are manufactures rather than retail consumers. Mr. Su said that we could try some marketing strategies such as exhibitions and seminars, it is important to let the target consumers know the advantages of the yeast we made.
Public Concerns
To obtain how the general public think about our project, we have released an online questionnaire survey, here is the core conclusions of the questionnaire survey.
Questionnaire
Picture 1
In order to gain a better understanding on people’s opinions about trans genetic yeast alcohol, our first survey was performed in forms of questionnaire. We gathered more than 500 samples in 3 days. The interviewees covered all age groups, from teenagers to seniors, most of them are aged between 31 to 45 years old. As shown in picture 1, about 80% of them use alcoholic products in their daily life, such as alcoholic beverages, hand sanitizer, cosmetics and ethanol-added gasoline. Which is quite a large amount of people. Based on their response, we can conclude that a number of people are purchasing alcoholic products, which lead to a huge demand of alcoholic products. This create a large market for the trans genetic yeast we are recently working on since manufactures are more willing to use a better, cheaper and more environmental-friendly yeast during the process of alcohol-producing.
Picture 2
In addition, as it shows in picture 2, only 13.80% of interviewees do not agree with the use of trans genetic yeast alcohol. We can draw the conclusion that after the alcohol produced by the genetically edited yeast enter the market, it won’t be opposed by most people, which confirm my previous conclusion. This also offers us determination and courage to undertake further experiments.
Picture 3
As shown in picture 3, when it comes to the alcoholic products people use, medical alcohol used for cleaning possess the largest utility (78.51%), while others are relatively average, which indicates the importance of alcoholic products in people’s daily life.
Picture 4
Furthermore, as picture 4 shows, most people have already be aware of difference the genetically edited yeast will make on the alcohol manufacturing industry. The result gave us a positive response, because at least 48.97% of interviewees are aware of the positive influence the trans genetic yeast have. As a result, we believe the genetically edited yeast will be supported.
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