Overview

Fig.1 Flow chart of E. coli carbon utilization system

1. Product design

In this project, we, the NCKU Tainan Team, have proposed an alternative way to reduce the emission of Carbon dioxide (CO₂). Referring to the opinions and feedback from many industry experts and professors, we design a new factory flow to capture CO₂ by E. coli. Not only our device meets the specs to commercialize, but it also demonstrates high cost performance.

The emission of CO₂ has been a serious problem for a century that causes global warming and severe climate change. Even though many ways have been tried to reduce it, the generation of CO₂ primarily from industry is still overwhelming. Therefore, scientists and governments have been working hard to find solutions to tackle the problem.

2. Enterprise interview

We complete the whole product design and propose to several industrial. Through the interviewing, we were able to stand in the site of customer and then improve our product. We found out what we were weak in the design and then figured out the solutions with the enterprise support.

1. Our major potential client China Steel Coporation, which has a large amount of CO₂ emission every year.
2. The national science research and development institute in Taiwan Industrial Technology Research Institute, which has the advance technology solving excess CO₂ emission problem.
3. An industry-university cooperative research project Biofixation　of　flue gas　CO₂ with microalgae , which is a relative project to us in Bio-method dealing CO₂ problem.
4. We collaborate with King Membrane Energy Technology Inc, which work for replacing traditional energy-using distillation with advanced energy-saving membrane technology.

Entrepreneurship

China Steel Corporation

Fig.2 Picture of CSC interview

Meeting with experts and stakeholders is important in shaping our project to fulfill the needs of our target user. China Steel Corporation is the largest integrated steel Manufacturer in Taiwan. Also, they had been adopting the algae bio-sequestration by cooperating with the research group at our university.

Process

We were given the opportunity to meet with the senior executive of China Steel Corporation to gain invaluable insight for our research. The meeting commenced with our presentation. During the presentation, we introduced our project, including the bioreactor design and the industrial model. By listing out all the aspects we had considered, we would like to obtain advice on the practical and social considerations involved in the application of our project in industry.

Suggestion and Question

Will the high concentration of CO₂ retard growth of engineered bacteria?

Microalgae is reported resistant to SOx and NOx. Does E. coli survive under such conditions?

The best condition for engineered E. coli to capture CO₂ is a lower CO₂ concentration without too much SOx and NOx particles. However, we won’t be able to provide an ideal culture condition in Industrial application. After testing the tolerance of E. coli, we conclude that E. coli is possible to survive under that kind of condition in factory and the only effects its expression. It may not capture as much CO₂ as culture in the lab.

It is important to define a specific commercial product that can be truly produced since your user may consider its economic viability. They stated that a product that can be widely used is better. At the same time, we should consider current GMO legislation if we want to commercialize those products. The actual condition is not as ideal as in the laboratory, we should optimize the condition to maximize the carbon fixation ability of the microbes.

Interview record

The record can be separated into two parts. One is about their feedback after interview, another one is our customer investigate questions. We use CSC represent China Steel.

Part1. Interview record

Date：September. 15, 9 am.

Location：China Steel Corp. conference hall

CSC： What is the adaptability of E. coli for the corporate? Do you have any doubt about the actual application?

It can be explained from the following points:

1. Concentration:

Bacteria can tolerate the increase of CO₂ concentration. However, there is limit in the input, and our team is targeting this system.

A shunt is designed to slow down the rate of input to enter the bacteria rapidly.

2. Temperature:

In this system, 42 degrees Celsius is our limit, and we need to overcome by technology in the high temperature.

The problem is that our team will lower the temperature through other devices.

3. Waste:

Our team solves the problem of waste by recycling and filtering out.

CSC ：From the perspective of the company, how much additional benefit can it bring to the output value of the products in their downstream of system?

At present, the product of downstream in our system is glutamine, why we choose is because glutamine is accessible and easy to operate for us. Its additional benefit refers to the different application. Take the market value of glutamine as example, the additional benefit can reach 10 times larger of the E. coli culture cost, ignoring the fixed cost of the whole system.

Besides, E. coli was regarded as high potential species to produce all kinds of protein. Including essential amino acid that cannot be synthesized by organism, or forage for stock farmer. Therefore, our system has high potential output value to bring great additional benefit.

CSC：China Steel is the second largest carbon consumer in our country. It needs two-thirds of Taiwan's area to balance one-tenth of the current emissions. In practice, it is still too far away. Is it possible to match the materials with 3D layout?

We want to save the space and culture in high density concentration:

1. Reduce the volume of culture material
2. Stacking the bioreactors

CSC： How to deal with the waste of this system? Is there a problem with super Cryptococcus neoformans?

The protein needs to be separated before produced. At the same time,this process will produce the bio-waste. The special process is high temperature and high pressure. It can be used in the factory's original waste system under the high temperature and high pressure environment.

We use the general strains, and there is no possibility of mutations. In addition, with the monitoring of environmental, the probability of mutation is greatly reduced to reach biosafety.

CSC position description：

Algae is one of the implementation of the CCS plan, and they always want to build a multi-system. Each system has its advantages and disadvantages. Therefore, what we proposed was a one more choice for them and they were glad to hear that E. coli and contribute to CCS&U (Carbon Capture Storage and Utilization).

Part2. Customer demand investigation

1. The research and development of new technologies, which level will be considered to mature and worthy investing specifically?

There are three conditions:

1) Feasibility of laboratory technology: It’s ok with technical confirmation.

2) Feasibility of engineering: It’s feasible under engineering equipment construction, application of space and on-site environmental conditions.

3) Feasibility of economic: total cost (input, output) must be positive benefits.

2. There is a problem of limited space in Taiwan, how much space did we need to reduce at least in the enterprise?

This proposition should be how much CO₂ the technology can absorb per unit area. Based on this basis, Industrial will evaluate the existing space of the factory, consider how much CO₂ can be absorbed, investment cost of equipment, the amount of CO₂ that can be reduced, and calculate the input and output to evaluate whether there is positive benefit.

3. We will consider the secondary cost of waste disposal, just like the application of your company unit in basic-oxygen-furnace slag, will you consider the cost of waste recycling be beneficial? Or is there a problem caused by China Steel and secondary pollution?

This part cannot be provided due to operational confidentiality. It is recommended that this proposition should be turned into be directly used as a marketable product. The cost of the resource should be assessed by the Life Cycle Assessment (LCA) as a whole.

4. Since our project is facing the problem about the higher cost of culture medium, we would like to ask you about the benefit of carbon fixation and cost of carbon fixation method.

The cost of carbon fixation depends on the carbon capture and storage methods used. For example, the calcium circuit developed by the Industrial Research Institute is used to capture carbon. The recent cost of carbon capture is intended to be reduced to US$30 per ton, and US$10 per ton of geological storage is required. Competition between carbon capture methods can be assessed by cost and overall utilization of reuse.

5. Regarding the part of industry-university cooperation, I would like to ask why China Steel chose to cooperate with Annan Campus in NCKU for microalgae carbon fixation.

When the former academic research unit strives for the NEP project (National Energy Program), the technology that the audited authority usually requires that project must be adopted by the industry. Therefore, both the academic research center and the industry usually sign the cooperation letter of intent for review. For China Steel, it is willing to support the academic research community to conduct forward-looking technical research with national resources to provide the technical information needed to evaluate feasibility.

6. The medium we need will still consume energy in the process of preparation, and it may cause carbon emissions simultaneously. We wonder how to regard upon overall carbon footprint may be increased from the perspective of enterprise.

If the overall footprint of the carbon fixation process developed may be positive (increased), in general, from the perspective of carbon reduction within the enterprise, there is no possibility of application. If the derived external carbon reduction benefit is greater than the internal carbon loss, it proves to have a positive net benefit to the environment. As long as it meets the feasibility of engineering and economic, the enterprise is willing to adopt it.

7. Research on carbon fixation, what is the driving force for China Steel in addition to economic benefits?

Regulatory requirements, corporate identity and social responsibility.

Part3. Picture Record

International Technology Research Institute

Fig.3 Picture of ITRI interview

Process

We attend the Biotechnology Green Energy Expo and have a business matchmaking with International Technology Research Institute (ITRI). The Dr. Cheng was a manager of Carbon Capture & Storage (CCS) Application Project in Green Energy and Environment Research Laboratories of ITRI. He gave a short speech about the condition of CCS in Taiwan. CCS technology is a relevant technology with our E. coli carbon utilization system. Therefore, through business matchmaking, we introduce our project and all the design including what we improve after the customer investigation with CSC.

Suggestion and Question

The currently solution in Taiwan is CCS, however, the policy and the inhabitant are big challenge of CCS technology. ITRI had cooperated with lots of industrial and academic institutes and developed advance CCS technology, however, most of them cannot have real implementation. The public doubt the safety about storage high density carbon underground. Therefore, ITRI now contribute to develop carbon utilize technology. That’s the point that ITRI will adopt our project into their lab.

Dr. Cheng suggested us to think more about the bioproduct after our E. coli uptake CO₂. We can easily transgene E. coli to let E. coli metabolized CO₂ and produce amino acid. The highest value of bioproduct will be the health food. However, we could hardly to make the health food which source came from industrial flue gas into the market. There are still many valuable product we can achieve with less limitation when apply to the market, such as electronic material solvent, biocytoculture, agricultural chemicals…...etc. Besides, the last choice will be biofuel. We should try to reuse our product again and again to maximize the value before converting it into biofuel.

This suggestion triggers our design about recycle system (link) after bioreactor.

Interview record

Part1. Interview record

Date：October. 6, 14 pm.

Location：Biotechnology Green Energy Expo conference hall

1. Is the bioreactor technology being involved in ITRI?

There is a biotechnology laboratory in ITRI. They work for convert microalgae into bio-fuel and dry anaerobic fermentation. Therefore, bioreactor technology was involved in ITRI.

2. After the carbon capture process, how to transfer those captured carbon dioxide while being condensed?

Once CO₂ was captured, we will liquefy it to decrease volume and then the vehicles will carry those high density liquefaction CO₂ to storage. Or through piping, we can transfer captured CO₂. to north of the sea for sequestration.

However, technology of carbon capture and storage (CCS) didn't implement in Taiwan, since that the inhabitant in Taiwan regarded CCS as environmentally hazardous technology. More and more national disaster due to climate led to local doubts about the safety of CCS. The currently policy of reduce carbon emission is to reach 2% decreasing CO₂ emission on 2020 compared with the standard year. The long-term emission reduction goal is to cut emission in 2050 by 50% compared to 2005 levels totally we hope to clean 1 hundred million tons of CO₂. The main challenge was the downstream process of CCS, utilizing technology.

According to the Greenhouse Gas Reduction and Management Act which was passed by Taiwan’s parliament (the Legislative Yuan), 1 tons of carbon tax is 100NT dollar, which is that industrial working hard to avoid high tax. However, what is the next step?

3. In the downstream process, can we reuse the waste heat produced from the factory for sterilization?

That is not a big problem. The temperature of waste heat produces from factory is around 100 to 1500 and 150 degree Celsius is suitable for our sterilization. So it is kind of a win-win situation because it do not cost extra energy or money for factory.

4. Taiwan does not focus on carbon sequestration, so will Taiwan use our technique? How to reduce the cost?

From the cost side, it is one thousand per gram because is not large scale but only laboratory level. How much cost can be reduced for one ton? How much cost can be reduced if we enlarge the scale? It does not cost too much reusing the industrial waste.

5. Now the technology renewable energy is mature, why not convert carbon dioxide into energy?

If we convert it to bio-fuel, the cost is NTD 50 (2 dollars) per litter. And the price that China Petroleum Corporation (CPC) sells is NTD 30 (1 dollar) per litter. Although the government will subsidize NTD 20 (1 dollar), it might not be a long term strategy. The industrial must find a way to reduce the cost itself. Besides, the edible product is more valuable than energy, but it also has higher limitation about listing.

Microalgae cultivation demonstration plant

Fig.4 Picture of An-nan interview

Process

In order to obtain more information about CO₂. biofixation, we visited microalgae cultivation in An-Nan Campus of National Cheng-Kung University which is a biofixation project managed by Professor Jo-Shu Chang. The research fellows showed us different scale of microalgae culture system, including open pond and photoreator. We also have a discussion about the position of biofixation in CO₂ emission problem. They shared their experience along the way developing the whole microalgae fixation system in ten years.

Suggestion and Question

Microalgae cultivation plant and our system both contribute to carbon utilization. The An-nan campus have advanced and well developed technology. Therefore, the visit was important to us when designing the whole E. coli carbon utilization system, especially the bioreactor design. Besides, we have discussion with each other to define the different advantages and disadvantages between different system. For example, the sunlight was a determine factor of microalgae while engineering E. coli. was not sensitive with light intensity. However, microalgae successfully converted CO₂ into valuable bio-product. Therefore, we concluded that different system will have different benefit depends on different demand.

Visit record

Date：August. 8, 10 am.

Location：National Cheng Kung University An-nan campus

1. Can the microalgae survive with flue gas from the industrial?

The flue gas from CSC contained about 6~7% CO₂ which is affordable for microalgae cultivation. Some species of microalgae can even survive under higher SOx and NOx condition. Therefore, the flue gas can be piped into microalgae cultivation demonstration plant directly without pre-adjusted.

2. How does microalgae culture control gas flow, temperature and pH?

With PE material, the air pipeline is plugged into the bottom of the culture pool. The pumped air can also help them stirring. Aspirator connected to a pore on the top to exhaust the net air. Since that the culture pool is implemented under the shade roof, temperature controller isn’t required. Besides, pH condition must be maintained around 7 while pre-culture.

3. Before culture the new species of microalgae, how to clean the pool completely?

First, exhaust the medium from the bottom and then jet the water to remove the microalgae attachment. Then, add some bleaches to kill the rest of microalgae in the pool.

Secondly, microalgae in the effluent medium will be separated by centrifugation. The separated microalgae will be storage through lyophilization. The microalgae attachment problem in the pipeline can be clean through jetting water and bleached as well.

4. What is the value of byproduct?

The separated microalgae can be used to reed shrimp since that the microalgae is one of the important nutrient source of shrimp. Through the shrimp farming, it can also extract Lutein which has high commercial value in the market.

5. How to select the specie of microalgae?

Before culture microalgae in the open pond, we will pre-culture in the lab and test under different conditions. Through many times experiments, we can select the final species of microalgae which has the best growth condition, likes heat-resisting.

Picture record

King Membrane Energy Technology Inc.

Fig.5 Picture of KME interview

Sewage problem is a critical issue for every kinds of bioreactor and ferment, especially for bioreactor containing genetic modified organism, which must absolutely prevent the organism leaking out and polluting the environment. After designing a bioreactor, we were eager to built up a sewage treatment system.

Process

On October 12, we visited the King Membrane Energy Technology company (KME), which is specialized in producing the Membrane Bio-reactor system. Different from traditional Sequencing Batch Reactor Activated Sludge Process, the system use hollow filter membrane that can filter most of bacteria.

Feedback

This technique minimized the area required for sewage treatment, allowed us to concentrate the medium before extracting bioproducts, and recycled the water after filtering. After the discussing with Dr. Kao, the manager of the company, we improved our device with the MBR system. This improvement made us one step closer to a company and eco-friendly bioreactor system.

Picture record

Business Model

The business model describes how an organization creates, delivers, and captures value in an economic, social, cultural, or other environment. Therefore, we introduce this business model as the basis for assessing the integrity and effectiveness of our ideas to work with our industry and even national research. First, we ask questions about this, and beyond the solution, we also explain why we chose this question. Second, we analyzed future developments, including the advantages of using this approach. Next, we introduce our plan to many relevant departments and discuss with the national research. I hope that this plan can be used to promote this plan in the future.

Target issue

More and more people are now paying attention to the impact of CO₂. The trend of environmental degradation is gradually increasing. Scientist and national worldwide contribute to capture those excessive CO₂. However, how to reduce carbon and use it has become a major problem today. Challenges against carbon process are complicate. Except the technique and implement problem, social acceptability and policy are other key factors about carbon process technology.

In general, planting is a method of carbon process, and the current use of green algae as a method of carbon utilization. This year, we hope to combine synthetic biology with the most advanced technologies. We want to draw people's attention to the environment and reuse these environmentally stimulating projects.

Business model analysis

Cost Evaluation

The cost evaluation is always crucial for product being on the market. To compare our engineered E. coli to microalgae, we calculate how much the cost it would be when capturing 1000 kilograms CO₂.

Volume

Table 1 Volume require in capturing 1000 kg CO₂

Cost

The most expensive source in the medium of our engineered E. coli is xylose. 1 mole xylose will capture 0.17 mole CO₂, so it would need 20.0535 kilograms xylose and 1 kilogram xylose is cost 2 USD. The total cost for our engineered E. coli is require 40.107 USD for capture 1 tons CO₂. In contrast, microalgae need 1000 liter to capture 250 gram CO₂, so it need 4000 liter (about 4 Tons) water and 1 tons is cost 9.78 USD (300NT). The total cost for microalgae is require 39.13 USD.

Table 2 Cost require in capturing 1000 kg CO₂

References

1. Fuyu G, Guoxia L, Xiaoyun Z, Jie Z, Zhen C and Yin L. Quantitative analysis of an engineered CO₂-fixing Escherichia coli reveals great potential of heterotrophic CO₂ fixation. Gong et al. Biotechnology for Biofuels, 2015, 8:86.
2. 張嘉修、陳俊延、林志生、楊勝仲、周德珍、郭子禎、顏宏偉、李澤民 (2015), 二氧化碳再利用─微藻養殖, 科學發展　2015 年 6 月│ 510 期
3. Lawrence Irlam (2017), GLOBAL COSTS OF CARBON CAPTURE AND STORAGE, Global CCS Institute, Senior Adviser Policy & Economics, Asia-Pacific Region
4. Jin Hwan Park, Jae Eun Oh, Kwang Ho Lee, Ji Young Kim, and Sang Yup Lee. Rational Design of Escherichia coli for L‑Isoleucine Production. [ACS Synth Biol.](https://www.ncbi.nlm.nih.gov/pubmed/23656230#) 2012
5. M. KUNDAK, L. LAZI], J. RNKO. CO₂ EMISSIONS IN THE STEEL INDUSTRY. METALURGIJA 48, 2009
6. V. N. Kalpana, D. Sathya Prabhu, S. Vinodhini and Devirajeswari V. Biomedical waste and its management. Journal of Chemical and Pharmaceutical Research, 2016
7. Qian Ma, Quanwei Zhang, Qingyang Xu, Chenglin Zhang, Yanjun Li, Xiaoguang Fan, Xixian Xie, Ning Chen. Systems metabolic engineering strategies for the production of amino acids. Synthetic and Systems Biotechnology 2 (2017)
8. Jørgen Barsett Magnus, Daniel Hollwedel, Marco Oldiges, and Ralf Takors. Monitoring and Modeling of the Reaction Dynamics in the Valine/Leucine Synthesis Pathway in Corynebacterium glutamicum. Biotechnol. Prog. 2006
9. Isao Kusumoto. Industrial Production of L-Glutamine. American Society for Nutritional Sciences, 2001