Team:SZU-China/Partnership
Partnership
Our project hit a bottleneck because the bio-indigo in the lab did not perform well. After we consulted a lot of literature without any results, we even thought about giving up.
We started to look for teams with similar projects, and learned from CCiC (Conference of China iGEMer Community) that part of the project of Shanghai_SFLS_SPBS used bio-indigo to dye hair. We contacted them and tried to seek help to solve our problem.
In order to troubleshoot our project, we carried out a deeper communication, sharing contacts, resources and materials without reservation. We had heated discussions about the application of bio-indigo. Although later in our project, we iterated to Gardenia Blue to achieve a greater variety of colors, we still helped each other a lot in experiments, human practices and hardware.
Fig 1. Logo of our partnership
Background Survey
Meeting–Get to know the projects of each teams
In order to have a better understanding of each teams' project, we held an online meeting. In the meeting, we talked about the impact of COVID-19 on our projects, how we tried to tackle these problems, introduced our projects, and discussed the production and application of bio-indigo. We also discussed whether bio-indigo is a good dye.
Background Learning–Learn more about dyes
Our projects are related to biological dyes, but we know little about biological dyes in the real market. Therefore, we decided to interview stakeholders to understand the scale and shortcomings of natural dyes in the current market.
SZU-China took a visit to a Weaving and Dyeing Company. They experienced the weaving and dyeing methods there, learning that they still need to spare no effort to reduce the pollution caused by dyeing processes.
They also went to the jeans washing water factory in Zhongshan city, Guangdong province, where they had a detailed conversation with the workers and leaders of the factory. It really shocked them that the workers work in such a poor environment that there was a pungent smell everywhere due to the volatilization of harmful chemicals when using chemical dyes. The team members couldn't stand it merely after ten minutes, while the workers need to work all day in such an environment. The factory managers introduced that it takes billions to deal with the water pollution caused by dyes. SZU-China, therefore, hopes to solve this problem in a more eco-friendly way such as using synthetic biology to produce a harmless dye.
They additionally talked to a hairdresser at Fazhixiu. They learned that customers are indeed concerned that hair dyes may damage the hair and scalp and cause health problems. Although natural plant hair dye products exist, the hairdresser admitted that synthetic hair dye products are cheaper. The hairdresser showed great interest when the team introduced their plan to produce natural pigments with engineered bacteria.
Finally, we shared the information, and reached a consensus to reduce pollution and other harms caused by chemical dyes.
Summary–Discussions in each team
After summarizing the various solutions that we had discussed, we tried to put them into practice. Shanghai_SFLS_SPBS decided to produce diverse biological dyes for hair dyeing. SZU-China decided to focus on the indigo dyeing problem in the jeans production and make efforts to improve the jeans production industry. After understanding the problems in the actual situation, both teams began to search literature to form a preliminary solution.
Experiment
Troubleshooting
We exchanged after completing our respective project designs, and we found that we will all characterize the same component–Synthesis of bio-indigo. We used the part of GreatBay_SZ (iGEM 2019), and tested the improved one of Shanghai_SFLS_SPBS. We found that the production of bio-indigo is difficult to further increase. In the experiment of Shanghai_SFLS_SPBS, the pET-28b(+) plasmid vector was used to increase the expression of TnaA and FMO enzymes. SZU-China tried to increase the tryptophan content in the medium to improve the expression of bio-indigo. But these attempts all point to the same result: no bio-indigo was produced, but instead the bacterial solution was orange-red.
We were all confused about this result, and therefore held a meeting to discuss this problem. We inferred that isatin may account for this color. When FMO is overexpressed, the produced bio-indigo is oxidized to be isatin, resulting in a decrease in indigo production, or none.
Having consulting a large number of documents, we decide to try transferring a glucosyltransferase gene to stabilize indoxyl, preventing spontaneous dimerization by protecting the reactive hydroxyl with a glucosyl group in the future.
Human Practices
Public Engagement
In order to verify the dyeing effect of our dyes, we organized the iDYE workshop. Shanghai_SFLS_SPBS provided us with dyes of various colors as a display, but considering safety issues, we did not directly use the dyes. We use food-grade gardenia blue for dyeing. The workshop attracted the attention of many students, and they created a lot of fascinating works!
Fig 2.Photos of the workshop
Questionnaire Survey
We conducted a questionnaire survey on biological dyes and synthetic biology. The two teams collected data separately, and then we conducted a unified summary analysis. Through analysis, we found that the public has a higher acceptance of biological dyes and generally believes that biological dyes are safer and more eco-friendly than chemical dyes. What's more, young people have a higher acceptance of synthetic biology. The acceptance of synthetic biology among older populations is lower, probably due to the more traditional point of view.
References
[1] Hsu, T., Welner, D., Russ, Z. et al. Employing a biochemical protecting group for a sustainable indigo dyeing strategy. Nat Chem Biol 14, 256–261 (2018). https://doi.org/10.1038/nchembio.2552
[2] LIU Hai-shun, ZHANG Zhen-wei, YANG Yu-ping, WU Xu, ZHANG Cun-lin. The Far Infrared Spectra of Five Dyes[J]. Spectroscopy and Spectral Analysis, 2018,38(10): 3064-3069. Doi:10.3964/j.issn.1000-0593(2018)10-3064-06
In order to have a better understanding of each teams' project, we held an online meeting. In the meeting, we talked about the impact of COVID-19 on our projects, how we tried to tackle these problems, introduced our projects, and discussed the production and application of bio-indigo. We also discussed whether bio-indigo is a good dye.
Our projects are related to biological dyes, but we know little about biological dyes in the real market. Therefore, we decided to interview stakeholders to understand the scale and shortcomings of natural dyes in the current market.
SZU-China took a visit to a Weaving and Dyeing Company. They experienced the weaving and dyeing methods there, learning that they still need to spare no effort to reduce the pollution caused by dyeing processes.
They also went to the jeans washing water factory in Zhongshan city, Guangdong province, where they had a detailed conversation with the workers and leaders of the factory. It really shocked them that the workers work in such a poor environment that there was a pungent smell everywhere due to the volatilization of harmful chemicals when using chemical dyes. The team members couldn't stand it merely after ten minutes, while the workers need to work all day in such an environment. The factory managers introduced that it takes billions to deal with the water pollution caused by dyes. SZU-China, therefore, hopes to solve this problem in a more eco-friendly way such as using synthetic biology to produce a harmless dye.
They additionally talked to a hairdresser at Fazhixiu. They learned that customers are indeed concerned that hair dyes may damage the hair and scalp and cause health problems. Although natural plant hair dye products exist, the hairdresser admitted that synthetic hair dye products are cheaper. The hairdresser showed great interest when the team introduced their plan to produce natural pigments with engineered bacteria.
Finally, we shared the information, and reached a consensus to reduce pollution and other harms caused by chemical dyes.
After summarizing the various solutions that we had discussed, we tried to put them into practice. Shanghai_SFLS_SPBS decided to produce diverse biological dyes for hair dyeing. SZU-China decided to focus on the indigo dyeing problem in the jeans production and make efforts to improve the jeans production industry. After understanding the problems in the actual situation, both teams began to search literature to form a preliminary solution.
We exchanged after completing our respective project designs, and we found that we will all characterize the same component–Synthesis of bio-indigo. We used the part of GreatBay_SZ (iGEM 2019), and tested the improved one of Shanghai_SFLS_SPBS. We found that the production of bio-indigo is difficult to further increase. In the experiment of Shanghai_SFLS_SPBS, the pET-28b(+) plasmid vector was used to increase the expression of TnaA and FMO enzymes. SZU-China tried to increase the tryptophan content in the medium to improve the expression of bio-indigo. But these attempts all point to the same result: no bio-indigo was produced, but instead the bacterial solution was orange-red.
We were all confused about this result, and therefore held a meeting to discuss this problem. We inferred that isatin may account for this color. When FMO is overexpressed, the produced bio-indigo is oxidized to be isatin, resulting in a decrease in indigo production, or none.
Having consulting a large number of documents, we decide to try transferring a glucosyltransferase gene to stabilize indoxyl, preventing spontaneous dimerization by protecting the reactive hydroxyl with a glucosyl group in the future.
In order to verify the dyeing effect of our dyes, we organized the iDYE workshop. Shanghai_SFLS_SPBS provided us with dyes of various colors as a display, but considering safety issues, we did not directly use the dyes. We use food-grade gardenia blue for dyeing. The workshop attracted the attention of many students, and they created a lot of fascinating works!
Fig 2.Photos of the workshop
We conducted a questionnaire survey on biological dyes and synthetic biology. The two teams collected data separately, and then we conducted a unified summary analysis. Through analysis, we found that the public has a higher acceptance of biological dyes and generally believes that biological dyes are safer and more eco-friendly than chemical dyes. What's more, young people have a higher acceptance of synthetic biology. The acceptance of synthetic biology among older populations is lower, probably due to the more traditional point of view.
[1] Hsu, T., Welner, D., Russ, Z. et al. Employing a biochemical protecting group for a sustainable indigo dyeing strategy. Nat Chem Biol 14, 256–261 (2018). https://doi.org/10.1038/nchembio.2552
[2] LIU Hai-shun, ZHANG Zhen-wei, YANG Yu-ping, WU Xu, ZHANG Cun-lin. The Far Infrared Spectra of Five Dyes[J]. Spectroscopy and Spectral Analysis, 2018,38(10): 3064-3069. Doi:10.3964/j.issn.1000-0593(2018)10-3064-06