From Bench To Business
Fig 1. Flow chart of E. coli carbon utilization system
- Product design
- Enterprise interview
- Our major potential customer, China Steel Corporation, which is the largest integrated steel maker in Taiwan. The iron making process involves reacting iron ore with a reducing agent, like coking coal, and produces large volumes of CO2.
- Taiwan's largest high-tech applied research institutions, Industrial Technology Research Institute, which has proposed several carbon capture technologies for use in industry.
- An industry-university cooperative research project ,Biofixation of flue gas CO2 with microalgae , worked on the same goal with us : reduce CO2 emission in industry with biology method.
- We collaborated with King Membrane Energy Technology Inc., which is a company that capable of providing customized pervaporation systems and membrane distillation systems for use in industries. They replace traditional energy-using distillation with advanced energy-saving membrane technology.
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 industry experts and professors, we design a new factory flow to capture CO2 by E. coli. Not only our device meets the specs to be commercialized, but it also demonstrates high cost performance.
The emission of CO2 has been a serious problem for centuries. The steep increase in atmospheric concentration of CO2 could potentially lead to a further increase in temperature and climatic change. Therefore, scientists and governments may need to take carbon capture technology to the next level.
We completed the whole product design and proposed to several industrial. Through the interview with our potential customers, we modified and improved our project from their perspective. We found out what we were weak in the design and then figured out the solutions with the enterprise support.
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 have been adopting the algae bio-sequestration by cooperating with the research group from our university, NCKU Tainan.
We met with the senior executive of China Steel Corporation to gain invaluable insight for our research. The interview started with our project presentation, including the introduction of 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 two questions above were the main concern of CSC. Basically, the best condition for engineered E. coli to capture CO2 have to be lower, without too much SOx and NOx particles. However, we won’t be able to provide an ideal culture condition in industrial application. After researching the tolerance of E. coli, we concluded that E. coli is possible to survive in factory condition while the concentration of SOx and NOx were much lower than CO2. Besides, we will dilute the concentration with gas that the small fraction of SOx NOx can only effect the expression of E. coli. In other words, 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 our 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.
The record can be separated into two parts. One is the feedback documentation during the interview, another one is customer investigate questions. We use CSC represent China Steel Corporation.
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:
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.
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.
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, and the reason 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 an 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 one of the largest carbon consumer in our country. In your presentation, you mentioned that two-thirds of Taiwan's area was required 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:
- Reduce the volume of culture material
- 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 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
- The research and development of new technologies, which level will be considered to mature and worthy investing specifically?
- There is a problem of limited space in Taiwan, how much space that we need to reduce at least in the enterprise?
- We will consider the secondary cost of waste disposal, just like the application of CSC 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?
- Since our project is facing the problem of the higher cost of culture medium, we would like to ask the question about the benefit of carbon fixation and cost of carbon fixation method.
- Regarding the part of industry-university cooperation, Why CSC chose to cooperate with Annan Campus in NCKU for microalgae carbon fixation.
- 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.
- Research on carbon fixation, what is the driving force for China Steel in addition to economic benefits?
There are three conditions:
1) Feasibility of laboratory technology: It’s okay 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.
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, and investment cost of equipment. Therefore, a certain amount of CO2 can be reduced. We will also calculate the input and output to evaluate if it has positive benefit.
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.
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.
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.
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.
Regulatory requirements, corporate identity and social responsibility.
Part3. Picture Record
International Technology Research Institute
Fig 3. Picture of ITRI interview
We attended the Biotechnology Green Energy Expo and had a business matchmaking with International Technology Research Institute (ITRI). The Dr. Shen 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 introduced 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 challenges 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. Shen 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 make the health food which made from industrial flue gas into the market. There are still many valuable products we can achieve with less limitation when applying 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.
Part1. Interview record
Date：October. 6, 14 pm.
Location：Biotechnology Green Energy Expo conference hall
- Is the bioreactor technology being involved in ITRI?
- After the carbon capture process, how to transfer those captured CO2 while being condensed?
- In the downstream process, can we reuse the waste heat produced from the factory for sterilization?
- Taiwan does not focus on carbon sequestration, so will Taiwan use our technique? How to reduce the cost?
- Now the technology renewable energy is mature, why not convert carbon dioxide into energy?
There is a biotechnology laboratory in ITRI. They work on converting microalgae into bio-fuel and dry anaerobic fermentation. Therefore, bioreactor technology was involved in ITRI.
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 ton of carbon tax is NTD 100 (35 USD), which is that industrial work hard to avoid high tax. However, what is the next step?
That is not a big problem. The temperature of waste heat produces from factory is around 100 to 1500 and 150 degrees 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.
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.
If we convert it to bio-fuel, the cost is NTD 50 (2 USD) per litter. And the price that China Petroleum Corporation (CPC) sells is NTD 30 (1 USD) per litter. Although the government will subsidize NTD 20 (1 USD), 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
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 photoreactor. 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 has 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 had a 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-products. Therefore, we concluded that different system will have different benefit depending on different demand.
Date：August. 8, 10 am.
Location：National Cheng Kung University An-nan campus
- Can the microalgae survive with flue gas from the industrial?
- How does microalgae culture control gas flow, temperature and pH?
- Before culture the new species of microalgae, how to clean the pool completely?
- What is the value of byproduct?
- How to select the specie of microalgae?
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.
With PE material, the air pipeline is plugged into the bottom of the culture pool. The pumped air can also help them stirr. 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.
First, exhaust the medium from the bottom and then jet the water to remove the microalgae attachment. Then, add some bleach 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 cleaned through jetting water and bleached as well.
The separated microalgae can be used to reed shrimp since that the microalgae is one of the important nutrient sources for shrimp. Through the shrimp farming, it can also extract Lutein which has high commercial value in the market.
Before culturing microalgae in the open pond, we will pre-culture in the lab and test it under different conditions. Through many experiments, we can select the final species of microalgae which has the best growth condition, like heat-resisting.
King Membrane Energy Technology Inc.
Fig 5. Picture of KME interview
Sewage problem is a critical issue for every kind 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 build up a sewage treatment system.
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 they use hollow filter membrane that can filter most of bacteria.
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 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.
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 analyze future developments, including the advantages of using this approach. Furthermore, we introduce our plan to many relevant departments and discuss with the national research. We hope that this plan can be used to promote this plan in the future.
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 have become a major problem today. Challenges against carbon process are complicate. Except the technique and implement problem, social acceptability and policy are aother key factors about carbon process technology.
In general, planting is a method of carbon process, and green algae is currently being one 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
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 1 ton of CO2.
Table 1 Volume required in utilizing 1 ton of CO2
|Organisms||CO2-utilization rate (mg/L*hr)||Biomass concentration (gDCW/L)||Specific CO2-utilization rate||Volume requiredd (L)|
|Engineered E. coli||19.6||0.87||22.5||51000|
Fig 6. Different volume required between micralgae and engineered E. coli
For capturing 1kg of CO2 in one hour, 51000 L is required with engineered E. coli carbon utilization. It seems that the difference volume required for utilizing same amount of CO2 is disadvantage of E. coli carbon utilization system. At this situation, we have to look into the design of the different bioreactor. For microalgae culture, it requires a large surface area to increase light intensity. As usual, the height of the microalgae culture pond cannot exceed 0.5 m. In other words, we have to build a 7 m diameter culture pond with the volume of 19000L. In constrast, engineered E. coli is not limited by light. The bioreactor of E. coli can be built with any height in the indoor or outdoor. To scale up the bioreactor, a 5.8 m diameted with 1.9 m height equals to 51000 L which has lower floor area required.
As a result,the bioreactor of engineered E. coli can save more than 30% floor area compared with micoralgae culture pond. Take the floor area of Taiwan as an example, we can build 94 billions of microalgae culture pond to uilize 10% of annual emission with 12 operation hours. However, 1 over 3 of floor area will be save if we replace them with E. coli bioreactor. E. coli bioreactor is more flexible on spacing using, and is less sensitive to weather effect.
The most expensive source in the medium of our engineered E. coli is xylose. 1 mole of xylose will capture 0.17 mole of CO2. Therefore, we need 20.0535 kg of xylose while 1 kg of xylose costs 2 USD. The total cost for our engineered E. coli requires 40.107 USD for capture 1 ton of CO2. In contrast, microalgae needs 1000 liter to capture 250 g of CO2, so it needs 4000 liter (about 4 tons) water while 1 ton costs 9.78 USD. The total cost for microalgae is 39.13 USD.
Table 2 Cost requireD in capturing 1 ton of CO2
|Item||Microalgae||Engineered E. coli|
|CO2 utilizing rate||250 g/m3/day||19.6 mg/g (DRY cell weight)|
|source required for 1 kg CO2 utilization||4 tons of water||20.0535 kg xylose|
|Cost||39.13 USD||40.107 USD|
|Source||NCKU Annan campus||Adjust reference and experiment|
We take two major industrial in Taiwan for example, which are China Steel Corporation (CSC) and Taiwan Semiconductor Manufacturing Company (TSMC). We had done some research on annual emission and calculated with our CO2 utilization efficiency. We also set the average carbon emission of small and medium enterprise (SME) as a standard goal which was easier to reach. Therefore, we can model the scale of E. coli carbon utilization system working for 1 % CO2 emission of different enterprise.
Table 3 Cost of dealing with 1 % amount of industrial CO2 emission
|Industrial||Annual emission||1 % of CO2 emission per hour||Number of required device||Area required||Operation cost (USD)|
|CSC||3.30 millon tons||3750 kg||4555||11.3875 hectare||150.4 thousands|
|TSMC||0.387 millon tons||442 kg||537||1.34 hectare||17.3 thousands|
|SME||20 thousands tons||23.529 kg||29||0.0713 hectare||1 thousands|
We take two major industrial in Taiwan for example, which are China Steel Corporation (CSC) and Taiwan Semiconductor Manufacturing Company (TSMC). We had research on annual emission and calculate with our CO2 utilization efficiency. Therefore, we can model the scale of E. coli carbon utilization system working for 1 % of industrial CO2 emission.
Our bioreactor applies in the industry, including the magnetic stirrer, pump and controller. It will cost 3313 USD every month if the price of industrial electricity is 0.063 USD per kWh.
Table 4 Energy consumption of different items of device
* hp = horse power
* kW = kilowatt
* kWh = kilowatt per hour in one month
- 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.
- 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
- Lawrence Irlam (2017), GLOBAL COSTS OF CARBON CAPTURE AND STORAGE, Global CCS Institute, Senior Adviser Policy & Economics, Asia-Pacific Region
- 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
- M. KUNDAK, L. LAZI], J. RNKO. CO2 Emissions in the Steel Idustry. Metalurgija4 8, 2009
- V. N. Kalpana, D. Sathya Prabhu, S. Vinodhini and Devirajeswari V. Biomedical waste and its management. Journal of Chemical and Pharmaceutical Research, 2016
- 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)
- 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
- Isao Kusumoto. Industrial Production of L-Glutamine. American Society for Nutritional Sciences, 2001