Human Practices | East_China

The ultimate goal of our group is to produce n-butanol through Lactobacillus “factory”, to improve the efficiency but reduce the cost of producing n-butanol, and reduce the pollution caused by the production of n-butanol in traditional methods.
Facing this topic, we also encountered many difficulties. For example, we didn’t possess enough knowledge about n-butanol in the beginning and most of people generally don’t know about n-butanol. Therefore, we tried to present the Abstract n-butanol into a lovely cartoon image in the charity sale part and the online social media posting to promote and publicize our project.
In the early stage of our research, we focused on figuring out the difference between our new method and the traditional method, as well as its market situation. Hence, we conducted a series of activities such as online questionnaires, street interviews to gather the opinions from public.
In addition, we managed to get in touch with BASF-YPC Company Limited which is quite famous for producing many raw chemical materials including n-butanol. As the important stakeholder in our project, their insights are valuable to us. Luckily we received the invitation to the field trip inside the company and had an interview with them after that.
By carrying forward the project, we realized that we need some professional perspective on what the issue is and how we could deal with it. Therefore, we contacted three experts in related fields and had virtual interviews with them, respectively.
This series of activities have made us break through ourselves again and again, and also increased our confidence in studying this topic. We hope that the topic we study can make some contributions to the world.

The purpose of this questionnaire was to investigate the public's understanding of n-butanol and pave the way for the future activities of our Buta Factory team. So far, we has received 164 valid questionnaires.
Below is the result analysis of each single question of our questionnaire.

As can be seen from the figure above, the main respondents who completed our questionnaire were teenagers under 18 years old and middle-aged people aged 40 ~ 50. To make the questionnaire more comprehensive, our team actively spread the questionnaire online and invited more people to take our survey. Because the elderly usually do not use smartphones, fewer questionnaires were collected over 60 years old.

Because n-butanol is mainly used in the chemical industry, we investigated people's impressions of chemical factories. It can be seen from the above figure that people are kine to believe that chemical production has brought serious environmental pollution (66.46%) rather than promoting the development of the economy (33.54%). But we still can not deny that the chemical industry does promote the development of society and it plays an important role in the social economy.

To understand the public's understanding of n-butanol, we designed this question. From this result, we can see that most people barely know n-butanol, no surprise. And a small number of people have only heard of the term. This tells us that we should make more effort to publicize this topic. So later on we decided to combine our topic with many illustrations, like our logo, to make them more understandable and acceptable to the public during our education part.

From figure 5, we can see that the main channel for people to gather information about n-butanol is the Internet. As a kind of raw chemical material, n-butanol seems far away from our daily life, so people will not take the initiative to learn about it. Most people who know about n-butanol due to the job regards. Therefore, our team thinks it is necessary to popularize some information about n-butanol to the public.

This question was designed to find out which aspects of n-butanol applications are of interest to the public. It can be seen that people are interested in many aspects of n-butanol, among which chemical production, fuel, and cosmetics are the most interested.

Currently, there are four methods of n-butanol production in the manufacturing industry and for most suppliers of n-butanol now the main method used in factories is carbonyl synthesis, which has high costs and unavoidable causes pollution to the environment. Most people feel that it is necessary to develop a new way to produce n-butanol. And our new solution to produce n-butanol with Lactobacillus could solve this problem.

According to our previous research, n-butanol can be used as a fuel. Thus we designed this question to investigate people's views on the prospect of n-butanol. Among them, 73.78% think there is a hope that n-butanol could be used to replace gasoline, while the rest think that this kind of transition will is very long and they are not optimistic.

Not only an irreplaceable material solvent in the chemical industry, but n-butanol has a broader application prospect in the market. We think it is feasible to popularize the use of n-butanol, such as replacing gasoline to be an energy resource. It can be seen from the above picture that most people support the development of n-butanol, which plays a supporting role in our research and proves that our project is responsible for society.

For this question, most people (74.39%) think that promoting n-butanol will be beneficial to environmental protection, which indicates that people pay more and more attention to environmental protection. Besides, many people (56.71%) also agree that this action will be beneficial to the local economy.

In order to further confirm the opinion of the public, our last question was set up as “one of two” between production efficiency and environment protection. Since our solution is still under development inside the laboratory it might not compete for production efficiency with the current methods of producing n-butanol in the manufacturing industry right now. But we believe that the traditional production of n-butanol, like other chemical raw materials, has inevitable pollution to the environment.
Based on our survey result, we have more faith to continue our project since for the majority (95.73%) environmental protection is the top priority, they think that we should focus on protecting the environment than improving production efficiency..

In the Popularizing Science Image Exhibition Hall of BASF-YPC Company, the commentator introduced the company's arduous entrepreneurial process, integrated mass production pattern, rich product system, and green and low-carbon environmental protection concept.
Historical photos and petrochemical products in the factory hall gave everyone a deeper and vivid picture in mind of the company's development history, general situation, and petroleum and petrochemical knowledge related to life.

During the visit, we enthusiastically asked questions and discussed them with the lecturer. After that, we had a deeper understanding of the company's product development and green environmental protection work. This valuable experience will give us deeper thinking about our project and how it could proceed.

Besides, we learned how they carry out green and safe production procedures on the spot and eliminated the misunderstanding of the chemical industry. We really appreciated them as this open form actively narrows the distance with students, so that everyone can improve their awareness of environmental protection while understanding petroleum and petrochemicals, and show the social responsibility and responsibility of this big enterprise.

In conclusion, even the traditional chemical production companies, are also seeking a greener, more eco-friendly, and more safe method or procedure to continue the production of these raw chemical materials. It is undeniable that the traditional chemical industry is facing a transition and they are looking for a transition. Our Buta Factory, aiming to produce n-butanol through biosynthesis and biotechnology, will have a promising future in this field.

The purpose of our Buta Factory research is to understand the knowledge of n-butanol in the general population.

Because of the epidemic, we were suggested not to go outside the campus, so we decided to conduct research inside the campus - Nanjing University of traditional Chinese medicine where we conducted our experiment as well.

In order to increase the chance of inviting people to take our survey, we took the initiative in restaurants, libraries, and other places to conduct the street interviews. Even if there was a risk of rejection, we still summoned up the courage to come forward one by one and asked patiently. “Hello, we are the Buta Factory team and we are doing social research. Do you have a minute for a quick interview? “

These seven questions are all our thoughts. The first question is to know the identity of t interviewee. The second question is to know the age of the interviewee. The third question is to understand the public's understanding of n-butanol. The fourth question is to understand the public's understanding of Lactobacillus. The fifth question is to understand the public's views on our project. The sixth is to find out which applications of n-butanol are of interest to the public. The seventh is to understand the public's views on the prospect of n-butanol as a fuel.

In the social research, most of the respondents are college students and most of them are medical majors. Among them, the majority of Pharmacists have only heard of the term but do not know much about n-butanol. We still need to strengthen propaganda and transform n-butanol, an abstract substance, into something easier to understand, which is convenient for people to know and understand.
In addition, The public feels that it takes a certain amount of time, technology, and, energy to develop new energy sources. But they still want cheaper and less polluting energy to replace oil. Therefore when introducing the wide applications of n-butanol in fuel, medical treatment, cosmetics, chemical production, food, and so on, they expressed their interest as well as support for the development of n-butanol. The public has great expectations for its application in various fields and hopes that it can develop better and better. This also gave us more motivation to continue this research.
Because our Buta Factory team is still exploring the biosynthesis of n-butanol by Lactobacillus fermenting (we believe that this method has lower cost, higher conversion efficiency, and lessens environmental pollution), we also interviewed the public about Lactobacillus. At least half of the respondents admitted that they didn't know about Lactobacillus, even not heard of it. Some of them added that they might see this term on the labels of yogurt products or other food labels, but not for sure. Therefore, it came to us that we shall also add the Lactobacillus to the educational topic list.
In general, we still need to make efforts to expand the public's understanding of n-butanol, Lactobacillus, and biosynthesis, including their explanation, their usage, their benefits, and harms, so that, at we really go hand in hand in a certain sense and further protect this beautiful planet together.

1.1 Competency analysis of production methods

Mr. Liu mentioned that there are three main production methods of n-butanol: propylene carbonyl synthesis method, acetaldehyde, or fermentation. The first method is adopted in China most, and the traditional fermentation method has certain problems:
1. Low yield and efficiency of producing butanol: Because of the toxic effect of butanol on bacteria, the mass concentration of butanol is less than 13g/L, the yield of butanol is less than 4.46s/(L·h), and the yield of butanol is less than 25%
2. Low final solvent concentration: The traditional ABE fermentation method has a solvent mass fraction of less than 2% and the water mass fraction can reach 98% or above, the conventional method increases the consumption of equipment, power, and energy, increasing the cost.
3. The proportion of butanol in the total solvent is low, generally accounting for only 60%, the remaining 30% is acetone, and 10% is ethanol, which increases the cost of butanol recovery and separation in the later stage.
4. Traditional butanol fermentation generally uses corn, honey, and other raw materials for production. With the rise of food prices and the shortage of world food resources, the development of butanol will be at a disadvantage.

1.2 Advice on improving our current solution

Mr. Liu believes that there are several ways to change the current problem:
1. Improve the existing strains: Use genetic engineering and metabolic engineering technology to improve the strain's tolerance to butanol, and then cut off the production and metabolism pathways of acetone and ethanol to increase the proportion of butanol in the solvent.
2. Discover more efficient and economically meaningful recovery methods, such as pervaporation, stripping, liquid-liquid extraction, etc.;
3. Optimize and commercialize the butanol fermentation process by using microorganisms and fermentation technology;
4. Expand fermentation raw material varieties, improve raw material pretreatment methods, and reduce the cost of butanol through systematic research.
On the last point, Mr. Liu added several plans in detail:
(1) Widely use cheap wood fiber materials that are easy to obtain such as straw, corn fiber, residual fruit of orchard, etc., because of lower cost, it has more commercial prospects. By 2010, China's rural straw output reached 726 million t/a, while the waste of the agricultural processing industry was up to 82 million tons. If these agricultural wastes are fully utilized, the yield of n-butanol can undoubtedly be improved.
(2) Study effective pretreatment methods to remove the toxicity of n-butanol through microorganisms and reduce costs
(3) Adopt a variety of fermentation methods to ensure microbial activity;
(4) Reduce the cost by coupling the steps in the bioreactor.
Solving just one of these problems would be a successful process Mr. Liu suggested.

1.3 Insights about the application of n-butanol

Mr. Liu said n-butanol is an organic compound with mild toxicity, commonly used as a solvent, or in the manufacture of paints, artificial fibers, detergents, etc. At present, N-butanol in China is mainly used for butyl acrylate, accounting for about 54%, followed by butyl acetate, accounting for about 18%.
In terms of properties, butanol has the following advantages compared with ethanol:
① High energy content, compared with ethanol can go 30% more distance
② Compared with ethanol, butanol has lower volatility, and a better phase than gasoline, but also has stronger adaptability
③ Butanol can be used in existing fuel supply and distribution systems, while ethanol needs to be transported by rail, ship, or truck;
(4) Compared with other biofuels, corrosion is less than ethanol, gasoline safety;
(5) Compared with existing biofuels, the mixture ratio of biobutanol and gasoline is higher, no need to modify the vehicle, and the economy of the hybrid fuel is higher;
Compared with ethanol, it can improve the fuel efficiency and mileage of vehicles;
Butanol produced by fermentation can reduce greenhouse gas emissions. As with ethanol, no SOx or NOx is produced when burned, which is beneficial to the environment.

Mr. Liu believes that if we successfully study the fermentation of lactobacillus to produce n-butanol, it can reduce the production cost or improve the production efficiency, and further develop the local economy. He shared some great cases with us.

Mr. Liu said that, in June 2006, Dupont of the United States and BP jointly announced the establishment of a partnership to jointly develop, produce and market a new generation of biofuel -- biobutanol, to meet the growing global fuel demand, the biobutanol plant will be put into operation in 2009. USDA Agricultural Research Institute (USDA-ARS) Research Program Cost-effective Bioprocess Technologies for Production of Biofuels from Lignocellulosic Biomass, Biobutanol is produced by conversion of cellulosic biomass by Clostridium difficile. The project was established in 2004 and is expected to be completed in 2009. In cooperation with EKB, GBL has invested 855,000 euros to innovate butanol fermentation technology. It plans to develop and produce butanol as a biofuel for transportation and reduce its production cost.Mr. Liu said that, in June 2006, Dupont of the United States and BP jointly announced the establishment of a partnership to jointly develop, produce and market a new generation of biofuel -- biobutanol, to meet the growing global fuel demand, the biobutanol plant will be put into operation in 2009. USDA Agricultural Research Institute (USDA-ARS) Research Program Cost-effective Bioprocess Technologies for Production of Biofuels from Lignocellulosic Biomass, Biobutanol is produced by conversion of cellulosic biomass by Clostridium difficile. The project was established in 2004 and is expected to be completed in 2009. In cooperation with EKB, GBL has invested 855,000 euros to innovate butanol fermentation technology. It plans to develop and produce butanol as a biofuel for transportation and reduce its production cost.

California Institute of Technology (Check), it's subsidiary Gevo, Khosla ventures, and VirginFuels have now switched from ethanol to butanol: Gevo will produce biobutanol from different types of biomass such as sugar cane, corn by-products, and grass. Enter Genetics International Inc. (EGI) uses DNA genetic improved strain, through metabolic engineering regulation and patent technology development of continuous immobilization reactor, using membrane technology to recover products, fermentation only 6h, the strain can tolerate 4%-5% butanol, Butanol in fermentation broth accounts for 90% of the total solvent (traditional fermentation butanol generally accounts for 60%), and the yield of butanol reaches 4.5-5.0g/(L·h) with a yield of 40%-50%, which is 400%-500% higher than that of traditional butanol process. The production cost is less than 0.264 USD/L, and the workshop cost is 5-10 million USD. The production cost of traditional acetone butanol fermentation is $2.50, and the traditional fermentation plant needs to invest at least $100 million.

Butyl Fuel uses Biobutanols patented by BFL. 1L corn can produce 0.27L butanol without ethanol or acetone. Currently, 1L corn can produce 0.14-0.20L butanol at most, and the ABE fermentation process is still used. According to preliminary cost estimates, the cost of producing butanol from petroleum is $1.350 / L, while the price of producing butanol from corn is $0.317 / L (excluding the hydrogen produced), which is competitive with the price of producing ethanol from corn at $0.338 / L. The cost of butanol can be reduced to $0.225 / L when corn is replaced with waste products such as feed. These companies; examples strongly prove that butanol has a bright future.

After the conversation with Mr. Liu, we were very pleased to see that currently biotechnology is gradually playing a role in the manufactury industry of raw chemical materials and we will join them to impact the world.

2.1 Feasibility analysis of our new production method

Prof. Wen said n-butanol fermentation in China is divided into three generations of the method, the first generation is to use the rest of the production of food to produce n-butanol, the cost is high, then the second generation of the production technology is to use cellulose degradation for sugar fermentation production, the cost is controlled in 8500 yuan/ton, but mainly with international oil production of butanol, the cost is controlled in 4800 yuan/ton, As oil prices soared and the price raised to 6,800 yuan per ton. In 2018, China also developed a third-generation technology to produce butanol, using factory exhaust gas, lowering the price to 4,200 yuan per ton. If all the world's butanol were produced using low carbon fermentation from factory exhaust, 16 million tons of CO2 could be saved each year. It is a feasible route to use Lactobacillus to produce n-butanol by fermentation. Traditional butanol fermentation has a big problem, that is, the toxicity of butanol is very strong, about 12g/L can kill microorganisms, but the skin of Lactobacillus is very thick, and can resist 30-40g/L butanol, so the upper limit of n-butanol production can be increased by using Lactobacillus. Therefore, Lactobacillus is regarded as the cell factory of the next generation of butanol, which has broad prospects.
Prof. Wen thinks that it is possible to put Lactobacillus in n-butanol industry production because the lactobacillus only faces a major problem, that is lactobacillus will produce more lactic acid, if you can overcome this problem, the lactobacillus to produce positive definite alcohol are very simple, lactobacillus industrial base is very good, so production will be very smooth. However, changing the habits of lactobacillus is a difficult job, because lactobacillus is prone to death, and it is difficult to achieve the desired effect.

2.2 Feasibility analysis of industrial production by the way of the “Buta Factory”

Prof. Wen thinks there are many uses of butanol, one of the biggest uses is used to do main chemicals. "Main" implies high demand, with 4 million tons of butanol needed annually, mainly for chemical production; The other thing is nonfungibility, ethanol, and other fuels can be replaced, but butanol is not.
Butanol is also a good fuel to use as an alternative to gasoline-gas-ethanol because it has very similar combustion properties to gasoline, and when the same amount of butanol and ethanol is added to an engine, butanol goes 30% longer and has fewer emissions than ethanol. Another important point is that butanol is a hydrocarbon and oxygen compound, which can be used to make hydrogen, and is 30% more hydrogen than ethanol. Moreover, China's west-east power transmission project will lose 20% to 30% of its electricity during transportation. It can use wind power and photovoltaic electricity to produce butanol, store the electricity in butanol, and then transport it to the east.
Prof. Wen believes that lactobacillus is the best choice for carrier selection.
American scientists have used Escherichia coli to produce n-butanol experiments, but failed, Escherichia coli is a high starting point, low upper limit, and lactobacillus is a low starting point, high upper limit. Escherichia coli is rich in tools and can be used in all kinds of operations, but its maximum yield is around 15g/L or even 30g/L, but it is difficult to industrialize, because butanol is toxic and risky, and factories are easy to lose money in production. Lactobacillus is less accident-prone, less risky, and more efficient
Prof. Wen believes that n-butanol has a very broad application prospect, the current market price is very high and the demand is also very large, as long as the initial lactobacillus can be developed for production, it can be put into production.

3.1 Insights about the application of n-butanol

Dr. Su believes that n-butanol is the most popular application as biofuel, and can also be used in medical, chemical, new energy, and other fields. However, Dr. Su believes that although n-butanol has a large demand, it still belongs to the range of chemical products that can be replaced.
According to Dr. Su, butanol is less volatile, corrosive, and dilutive than ethanol. When used as a fuel, butanol has more energy because it contains more carbon and has a lower boiling point than ethanol, so it burns for longer. Dr. Su gave an example: Ethanol has been mature in the United States as a fuel to replace oil, while butanol has higher combustion efficiency. In the future, butanol is likely to further replace ethanol and oil, which have great potential, but it will be a long process. Dr. Su gives the hydrogen-fueled car as an example, although hydrogen cars have many advantages, because hydrogen stations are not popular, coupled with some production costs, hydrogen cars have not been very smooth and have low development.

3.2 Advantage analysis of our solution compared to the traditional methods

Dr. Su thinks that using bacteria to synthesize n-butanol is a relatively advanced technology. At present, the main problem is how to reduce the cost while improving the yield, and remove some of its limitations or improve its metabolism through genetic engineering, to reduce the cost of separating n-butanol. Compared with the traditional petrochemical production of n-butanol, the petrochemical industry has developed nearly 100 years of history and is undoubtedly a very mature industry, so the cost is lower, but for the environment, the petrochemical industry will produce a lot of pollutants. By lactobacillus to produce n-butyl alcohol has first security is higher, because manufacturing n-butyl alcohol by biosynthesis, ruled out a large part of the safety problems, the second made by the method of biosynthesis of n-butyl alcohol, do not contain carcinogenic substance, so can be put into food, medical and other fields, but on the other hand, the lactobacillus has many advantages, However, due to the raw material acquisition and petrochemical industry is relatively difficult, so the cost of the price will be higher, in addition, because of the insufficient performance of the strain, it is likely to lead to the genetic transformation of the strain is difficult, and will produce a large number of by-products, reduce the utilization efficiency of the strain.
Dr. Su believes that the use of Lactobacillus to produce n-butanol has a very bright prospect as long as the problems of yield and raw materials can be solved.

After interviewing these experts, we had a clear picture of the current market and had a deeper thought about our current project design. They brought us not only professional insights as well as advice in which we could complete our business plan, modify the further research direction and improve our future development plan, but also the faith and support we could continue this project.