Welcome to iGEM Aachen 2016

Public Engagement

“Education is a common task of science and economy [...].”

Dr. Karsten Schürrle, DECHEMA

Establishing a dialogue with the public and getting them in contact with synthetic biology was something we feel is extremely important. Listed below, you can find in which ways attempted to spread the iGEM spirit.

Spreading the News

We wished to get in touch with as many people as possible. Hence, for presenting our work without us being physically there, we developed several ways to spread information about our project and the idea of synthetic biology.

Newsletter
From the beginning on, we received a lot of support from various sides and many people were interested in our project. That is why we thought of a way to inform them all at the same time about our progress and developed a monthly newsletter. It was sent to all of our supporters and everybody who opted in. With the newsletter, we intended to explain our project to people with and without the scientific background and therefore developed a short and an extended version. Writing it helped us reflecting about our work and also preparing for the wiki. We even received suggestions from experts who read about our main challenges in the newsletter.

Flyer Distribution
Our flyer was created in the beginning of our project to give interested supporters a short overview about our idea and to inform people about what we are doing throughout this year of participating in iGEM.
As iGEM is a great opportunity and experience for everybody - not only university students - and as we could not visit as many schools as we wished, we also sent an adapted version of the flyer with additional information about the competition to about 300 different schools.

Presenting iGEM and Our Project at Schools

Figure 1: "FCG" school in Düsseldorf.

Our team visited many different schools in and close to Aachen. To give the pupils a better insight in the field of synthetic biology, we prepared project days in cooperation with the teachers. Our overall goal was to give them an understanding of synthetic biology and microbiology and introduce iGEM to them.

We gave a presentation about synthetic biology, our project and the iGEM competition in general and afterwards gave a safety instruction and performed some experiments with them. During the visits, we were happy to answer all of their upcoming questions as they got more and more in touch with synthetic biology, our project and iGEM.

Since we visited different grades with students of different ages, we adapted every presentation and each experiment to the particular school and their demands. After the experiments and the dialogue with the students we spend some time with the teachers to reflect on our visit. While talking to them we discovered the lack of possibilities for them to perform biological experiments in schools. To meet those needs, we created a collection of easy to understand experiments that do not require any specific equipment.

Figure 2: Two team members at "St. Ursula" school, Monschau.

Book of experiments (To view the pdf-file, please click here)

Monschau
We visited a girl’s school in Monschau on two days. First, we talked to several teachers to plan and prepare our visit. We benefit from the teacher’s hints about which topics might be interesting for the students. It was a great pedagogical experience for us, as we had to think of a way how to explain the biological topics to students who do not have a huge biological knowledge.
We thought a lot about it and in the end, it was a great success. We could evoke curiosity and the girls asked a lot of questions about Synthetic Biology and iGEM.

Aachen
Moreover, we visited the St. Ursula Gymnasium in Aachen and presented our work to students who already had a good biological background through their school. We created a new presentation for this purpose and experiments adjusted to their level of knowledge. It turned out that many students were interested in choosing a biology-related topic as their study field. This led us to explaining more about our university and talking about different subjects which are interesting for example Synthetic Biology. Later, we had a lively talk with the teachers who gave us some feedback and discussed the role of genetic engineering in Germany.

Düsseldorf
In Düsseldorf, in cooperation with two different schools, we organized a whole “Biotech-Day”. It was an and challenging experience for us because there were 89 students for whom we needed to do a lot of preparation. But luckily, with the help of the teachers we managed to plan the day. We did not only present the theoretical background of our project, but also put emphasis on safety aspects related to synthetic biology and the experiments. Hence, we were able to initiate interesting conversations and discussed both, our project idea and the practical work in the lab.
As we prepared some long term experiments, we visited the school again after 2 days and could show the results to them. On that day, they took the chance to ask more specific questions about about Synthetic Biology and our field of study. What looked like an overwhelming organizational challenge turned out to be a great experience with students not much younger than us.

Postcard Project

This year the iGEM Team Düsseldorf started a postcard project. Every participating team was encouraged to design their own, with the purpose to increase the knowledge about synthetic biology in the public. We thought it was a great idea, so we participated. These cards have first been distributed amongst all participating teams and then were distributed in different locations around each team’s hometown.

With our topic we wanted to inform people about synthetic biology and genetic engineering and show that they are present in their daily life, in an easily understandable way.
Here, we want to thank once again the iGEM Team Düsseldorf for this great idea and the invitation to participate.

Click on the postcard to see the back.

English translation:

"What does synthetic biology even mean?
The term “genetic engineering” is often mentioned in critical media reports and in this context cheese manufacturers more and more advertise with the promise that their products were made without this allegedly dangerous technology. We, as students of this field, would like to give you a better understanding of synthetic biology with the following example: Nowadays, you can find the declaration “without gene technology” on cheese packages. This does not refer to the actual ingredients of the product, but to the production method, so the cheese itself does not differ from “normal” one. Synthetic biology makes it possible to generate products with the help of genetically engineered microorganisms. In contrast, cheese without genetic engineering is produced with dried calf stomachs, which raises the question if this is more desirable.”

Inspiring the Next Generation

Figure 3: Visitors from "Jugend forscht" in Aachen.

“Jugend forscht” is a European science and technology competition for pupils, with more than 10,000 participants annually. The pupils work on a self-chosen project and present their results at regional levels and later at a national contest to an expert jury. Some winners of this competition were invited to Aachen and visited different scientific places. We organized a presentation and a tour through the laboratory for them. It was inspiring to see, what an immense interested and advanced knowledge about science they have already have.
After the presentation, we had a chance to talk about our project and the iGEM competition.
It was amazing to be able to enthuse these promising young people further for synthetic biology.

Writing Competition

In the German society knowledge about synthetic biology is not very widespread: we found out that the possibilities for people to inform themselves about synthetic biology are very limited which causes a mostly negative attitude towards this subject. Many are sceptical and focused more on the dangers than the possibilities. That’s why we wanted to give especially young people an exciting opportunity to get in touch with synthetic biology via a writing and arts competition - submissions of all group of ages were welcomed. Participants were encouraged to deal with aspects of synthetic biology in a creative way and express how they feel about it. Combining their great ideas with this topic led to amazing stories, which built a bridge between the scientific society and open minded people who were inspired to create their own opinion about synthetic biology.
The winner in our iGEM writing competition 2016 was Hannah Lanzrath with her story titled “14. Oktober 2044”.

Figure 4: Prize for the best story.

Here you can see a short abstract:

17. Oktober 2044
“…Einatmen. Ausatmen. Konzentrieren. Anna versuchte ihre Gedanken zu sammeln. Jedes Detail könnte wichtig, könnte hilfreich sein doch noch abzuwenden was unabwendbar schien. Zweimal positionierte Sie den Kugelschreiber in ihrer schweißnassen Hand neu, bevor sie die Kugel auf das Papier drückte. August – 2034 Damals regnerischer Sommer. Globale Erwärmung sorgt seit 1980 für stetigen Anstieg in Anzahl der Regentage und Niederschlagsmenge pro Jahr. Getreide und Gemüsefelder überwässert, 30% der Ernte fällt aus. Folge: Massenimport von Getreide aus China. Dort seit 15 Jahren etablierter Getreideanbau auf neu erschlossenen Inseln. Frage: Sporisorium Lamia beim Import eingetragen?
Sie blickte von dem Notizblock auf. Wortlos hatte ihr ein hagerer, erschöpft aussehender Mann einen Zeitungsartikel zugeschoben. „Noch mehr?“ brachte sie mit zusammengepressten Lippen hervor. ...“

“…Breath in. Breath out. Concentrate. Anna tried to gather her thoughts. Every detail could be important, helpful to turn away what seemed inevitable. Twice she tried to reposition the pen in her sweaty palms, before she pressed the ball onto the paper. August-2034 Rainy summer. Since 1980, global warming effects steady increase of days of rain annual amount of precipitation. Corn and vegetables fields flooded, 30% crop failure. Consequence: Mass import of corn from China. Since 15 years Established grain productions there on newly accessible islands. Question: Sporisorium Lamia instroduced via import?
She looked up from her notepad. Wordless, a gaunty exhaused looking man pushed a news paper article towards her. „Even more?“ she moaned between her teeth….“

We want to thank again all the participants of the competition. We hope you had as much fun as we had.

Beyond The Basics

Apart from basic lab work we also established a dialogue with the public.
Our outreach included contributing to conferences, socializing and scientific events.
Beyond that we asked ourselves how to maximize the future impact of our project and made an effort to get as much feedback from outsiders as possible. This motivated us to evaluated the economic perspectives of our LIPs, as well.

Economical Perspective

Many iGEM teams create great ideas and great projects, but when it comes to the implementation and application in real life, it is also important to deal with the following questions: Would this project be applicable in reality? Would there be a market for this product? What opportunities are there to make the product more appealing to consumers? Apart from "Integrated Practices", it was also of great interest for us to examine the potential of our idea on the laundry detergents market.

That is why we created an economical analysis in cooperation with a business administration student from the University in Nantes, France.

Politics of the new product
The iGEM team Aachen wants to produce the photocaged Subtilisin E and the organisms, which can produce the new enzymes. So they are in direct competition to companies like Novozymes or Roche, which offer enzymes for different applications. Until now they only produce in a very small scale. Maybe they can work together with other enzyme producing companies or directly with washing companies. It would definitely be necessary to have an industrial partner to benefit from their experiences on the global market, because the German laundry detergent market is too competitive and controlled by a few corporations.

Politic regarding the distribution of the new product
The distribution of the new product depends on the way the iGEM Team Aachen wants to introduce the new product on the market. On the one hand they can try to industrialize it with the help of already existing enzyme companies. One the other hand their product could be applied in the washing industry and used for washing detergents. That’s the most probable application of their protease. With the help of these already existing companies it is possible to distribute their product. For example it could be sold in local supermarkets and drugstores. Probably it won´t be sold in low budget supermarkets, because of the production and developmental costs. Moreover, the iGEM Team Aachen has to care about the distribution of their light tool. Maybe they can include it as a free give-away to the first bottle of washing detergent and start in this way promoting the use of its product.

Politics of communication of the new product
The new product of the iGEM Team Aachen should be communicated as an alternative to already existing washing detergents and there should be a focus on the ecological impacts of boric acid. The product of the iGEM Team Aachen should ban boric acid and make it possible to use the washing detergents with the same quality as before. Instead of boric acid they want to communicate the use of light to activate the protease. As the iGEM Team wants people to activate the protease with light, they also have to communicate this difference. That’s where I see the biggest problem for the iGEM Team Aachen. They have to convince the people that they should not only invest more time in the washing process, but also to change their customs when it comes to cleaning their clothes. Moreover they would need to convince big companies. Although the iGEM Team invented a gadget for activating the protease and in a survey the team performed the majority of the people said that they would spend more time for activating the protease, this idea has to be transferred in the daily life of the customers. For that reason this device has to be as user friendly as possible, the application should only differ in a very small way from today’s application and the price should be in the same range like before. If they take care of these purposes and still maintain the ecological part, the products can have a chance on the global market

Special Events

NRW Day

Figure 1: Presenting our project in Düsseldorf

This year the state North-Rhine Westphalia celebrated its 70th birthday, with event days for the public, in its capital Düsseldorf. Because of that, the iGEM Team Düsseldorf invited members of our team and other iGEM teams from North Rhine Westphalia to come to Düsseldorf. Here we were given the possibility to present our project idea to the public and to talk about the advantages of synthetic biology. This gave us the chance to review our project and integrate the opinions, fears and ideas from other people. We want to thank the iGEM team Düsseldorf for organizing this.

Future Lab Gala

Future lab is a gala organized by different institutes and organizations in Aachen to present Aachen as the perfect place for research, science and industry. During this gala, the participants presented themselves with the help of scientific experiments, art, and speeches. It was a great opportunity for us to get in touch with politicians, people from the industry, and different universities and to tell them about iGEM and our project.

Figure 2: Our Team visiting the Future Lab Gala in Aachen

Conferences

Annual meeting of DECHEMA

Figure 3: Presenting our project at DECHEMA convention

In September, the team from Göttingen and us, the iGEM Team Aachen, were invited to the “DECHEMA Jahrestagung”. The “Society for Chemical Engineering and Biotechnology” gave us a slot to present our project in front of a scientific audience. Our presentations were included in the “Zukunftsforum”, a subitem of the conference with focus on the future perspective and strategies for the biotechnological industry in Germany. We are thankful for the opportunity to establish a dialogue with the scientific community and practice our presentational skills for the Giant Jamboree.

Analytica

We visited the trade fair “Analytica”, for analysis, biotechnology, laboratory technology, and devices in Munich. Here we got in touch with different companies and presented our project to them. Moreover, this trade fair gave us the possibility to win over supporters for our project. We also met members from the iGEM Team Munich.

SynBioBeta

SynBioBeta is one of the main communities of entrepreneurs, investors, and policy makers devoted to promote the responsible growth of the synthetic biology field. On their international conferences and events many members of the synthetic biology community come together and discuss possibilities to promote the development of this industrial field. The background of people, visiting these conferences, is very diverse and therefore various aspects are discussed. Our members were not only able to meet industrial members and scientist, but also other iGEM Teams and Randy, founder of the iGEM organization. This was an amazing experience for us and left a big impression on our members during their visit in London. Furthermore, our members were able to listen to different lectures and participate in many discussion rounds.

Figure 4: Some members of our team meeting Randy and other iGEM Teams at SynBioBeta

Biomedica

In 2016 the Biomedica trade fair took place in Aachen from the 30th to the 31st of May. So, many members of our team decided to visit this conference and look at the different topics, which were presented. Moreover, the four headwords: “Pharma, Biotech, Meditech, and Innovation” called our attention. Biomedica started in 2007, as the first Euregional conference on lifesciences. Biomedica started as a joint conference of three already existing conferences in Liege, Aachen and Maastricht. It focusses on combining business and science.

Meet ups

European iGEM Meet up

The iGEM Teams from Paris invited us to visit the European iGEM Meet up in Paris. For our members it was a great experience to participate, because they had the opportunity to get in touch with other iGEM teams, learn something about the different projects and establish a personal contact with other members. Moreover, they had the chance to include the expertise and suggestions from other teams in the planning process of our project. But of course we also had a lot of fun in this amazing city. In this context we want to thank once again the iGEM Teams in Paris for their invitation.

Figure 5: iGEM team gathering

German iGEM Meet up

Figure 5: Members of our team playing bubble soccer at the German iGEM Meetup

All German teams came together at this Meet up in Marburg. iGEM Marburg organized a tour through their town and for the scientific exchange, they invited local scientists, industry representatives, and of course Traci from iGEM headquarters to give talks. Moreover, we had the opportunity to present our project ideas in front of all the other teams. It was a lot of fun and after the weekend we went home tired but happy.

iGEM Team Göttingen

When iGEM Göttingen came to Aachen for DECHEMA we took the opportunity to show them around our city and have a nice time with fellow iGEMers.

iGEM Team Bielefeld

We visited Bielefeld and its university and with a nice barbecue and interesting chats, we enjoyed the social site of iGEM there.

Competitors and Partners at the Same Time

iGEM – an international competition in synthetic biology. Teams from all over the world are competing to win the Grand Prize at the finale in Boston. However, we did not only see each other as competitors, we also took the opportunity to get in contact with them and share our knowledge and experience to improve each other’s projects.

It is great to be part of the iGEM community and here we would like to thank every team who helped us in making our project in and outside the lab successful!

Click on the logos to find out more about our mutual support!

NCTU Formosa We found out that our universities are partners so it was obvious to get in contact via skype. We shared our knowledge concerning research and lab work. The Team from Taiwan also participated in our international survey and helped distributing it to other teams as well	XMU China The iGEM Team from the Xiamen University in China collected newsletters from international iGEM teams. We were happy to not only share our newsletter on a national basis but also international with their help.
TecCEM We got in touch with the iGEM team from Mexico. First we only helped sharing their survey. After that we stayed in touch with the team and further cooperation possibilities emerged. For example, they participated in our international survey and advertised it to other teams. Moreover, we participated in their international video project and made two videos in German for them.
Göttingen iGEM Göttingen visited Aachen and spend some time with us, when the DECHEMA Jahrestagung took place. While giving them a tour around Aachen, we were talking about the progress of our projects.	HHU Düsseldorf We participated in the postcard projec which they initiated. Moreover, we visited the NRW Day in Düsseldorf and presented our work to the public and to other iGEM Teams from North Rhine-Westphalia.

Contribution to METU HS Ankara Lab Protocols Collection

We were contacted by the iGEM Team METU HS Ankara. They had translated iGEM experiment protocols to Turkish and now had the idea to create a database consisting of protocols translated to different languages. We liked the idea of making those protocols accessible in every country, so we also contributed to this collection.

Thank you, for contacting us, iGEM METU HS Ankara and giving us the chance to add to the success of your project.
Click here to see the protocols we translated to German.

Survey

A useful tool to get different opinions on topics related to the iGEM team’s projects is conducting surveys. Therefore, you need as many participants as possible to get a broad spectrum of opinions. So our team decided to take part in the surveys of the different teams which can be seen below.
Thank you

Politics

We got in contact with many different politicians and asked them questions related to genetic engineering and the political situation in Germany.

The answers from the different political parties can be seen in the PDF files below.

Dr.Zerlein
Mr. Adenauer
Die Linke
Mrs. Schulze
Piraten Aachen

Ethics

Until now we did not look at the ethical perspective of our project. After our conversations with the public and some other professors it is not surprising, that synthetic biology, which adapts and designs living organisms with many different applications, has a broad ethical dimension. An ethical dimension, an ethical discussion … ethical questions … ethics?

Many of us maybe heard about ethics, because of their philosophy courses or because of their religious education at school. Some of us maybe saw an ethic expert in television or in some TV discussion rounds. But what is ethic exactly?

Ethic is a branch of philosophy and refers to well-founded standards of right and wrong. Normally it defines what humans ought to do and what is wrong about their actions. Ethics is about trying to give answers to questions about how we have to react to or act in specific situations, what we have to consider to develop a well- founded decision or what is wrong about the way we handle different topics.

As we do not know so many aspects related to ethics and because of our non-objective perspective, we started to contact different experts in Germany. The pdf-file of one of the interviews can be found below.

Dr.Ach

Perspective of Professors

Interview with Our Supervisor Prof. Schwaneberg

How do You evaluate the iGEM competition in general and the influence which the former iGEM teams from Aachen had?
In general, I see the iGEM competition as a very good opportunity for students to learn how to work and act in a team by proposing ideas, fighting for them, getting exposed to their consequences. In essence, it is a very good training for students to learn how to take over responsibilities and to work in and as a team. I strongly support the participation of students in the iGEM competition and it is great if they have the chance to travel to Boston and present their concept at the Giant Jamboree. There they will be exposed to other brilliant students all over the world which share common interests. Apart from this, the iGEM competition has a very positive role in terms of visibility for the nature science faculty of the RWTH Aachen University and even generated interest from companies.

How do You think: Can iGEM support the education of the students?
I would say that iGEM also has a very positive influence and shapes their personality to become leaders in research. Members of the iGEM team will get a totally different insight of lab work and research by developing own ideas and experience how challenging the realization on the molecular level can be. However, taking and learning from these frustrations enables students to grow by discovering and analyzing challenges as well as finally solving them totally on their own. The letter fosters their believes in own abilities and strengthens their personality and leadership skills to tackle important scientific questions in the future.

What makes iGEM special in comparison to other student programs and practical courses?
I think the most important part of iGEM is the aspect of self-organization in terms of selected topics and fundraising to support iGEM activities. The latter experiences makes iGEM outstanding and much more challenging than other student activities and internships in which research topics are usually provided by the supervisor.

How do You evaluate the complexity of the iGEM competition (financial support, organisation, and public relations)?
iGEM teams have more or less to act like a biotech start up company which has to develop and to market their own development to finance their challenging activities and thereby establishing their organization, public relations and fundraising. The complexity of the iGEM competition and the limited time for idea development makes iGEM a highly complex and challenging endeavor.

What do You think about this years iGEM team Aachen and its project ideas?
In one word: GREAT. Light switchable proteins are a very promising and challenging topic in biotechnology and in material science and will emerge in the future as a hot topic in national research agenda on bioeconomy in Germany.

Please name three terms that describe the iGEM teams in Aachen and their work?
The three terms that come directly to my mind to describe RWTH’s iGEM team Aachen are “international”, ”interdisciplinary” and ” innovative” (triple i work).

Could You please tell us something about the previous work of the iGEM teams in Aachen?
The past two iGEM teams from Aachen were SENSATIONAL. They won several gold medals and awards as best newcomers. I am very proud of our students!

How do You feel to be a part of the iGEM team Aachen 2016?
It is fun to discuss with them and it is my pleasure to support these excellent young scientists with experience/infrastructure and to see them to grow with their challenges and success.

Interview with Our Supervisor Prof. Blank

How do You evaluate the iGEM competition in general and the influence which the former iGEM teams from Aachen had?
The iGEM competition allows self-organized research. The aspect of bringing engineering into biology is fitting very well to the RWTH Aachen University and the Jülich Research Center. The three teams competing as the iGEM teams Aachen paved the way for synthetic biology at RWTH Aachen.

How do You think: Can iGEM support the education of the students?
iGEM can support the education because the students work self-motivated. Moreover the concept of research-based teaching intensifies the teaching experience. That the newest synthetic biology technologies can be applied is lifting the teaching in Aachen on a different level.. After their iGEM time many members have accelerate expertise and working skills. For sure, the softskills were practiced, working in a group of about 15 people. Students have the chance to develop new skills, new management experiences and to work with new devices.

What makes iGEM special in comparison to other student programs and practical courses?
A huge difference is that students have the chance to work on their own. Moreover, they have the chance to develop a project on their own and work on it till the end of the competition. Furthermore the students can get in touch with local companies and learn how to ask for financial supports for their project. Especially in Aachen, the topic is chosen by the students hence the students have the chance to integrate project ideas, in which they are really interested. Students have the chance to learn everything on their own and make special experiences, which they could not get from other student initiatives or other practical courses. These differences make iGEM clearly a unique experience.

How do You evaluate the complexity of the iGEM competition (financial support, organisation, and public relations)?
Again very positive. Students have the chance to integrate their different interests and talents into the competition and into the project. Moreover it is great that students can work several months together on the same topic and work with students who are interested in very different subjects. Additionally, this is a good practice for their professional life.
Attracting funds and raising public awareness for the project is central to iGEM. Without the initiative of the students the presence of iGEM in Aachen and the special iGEM spirit would not be the same. The RWTH Aachen University and the FZJ support the iGEM team every year. The supporters of the iGEM team think that the “Bottom up” strategy, i.e., students first, fits best to Aachen. Because the students have the chance to develop their OWN project, obviously with support from the experts and institutes.

What do You think about this year’s iGEM team Aachen and its project ideas?
Super, you can talk about the topic with many different people, who are unfamiliar with synthetic biology and biotechnology. Everybody uses liquid washing detergents because of the absence of dusts and allergenic substances. Moreover, it is possible to explain the environmental problem caused by boric acid to the public and still present a very interesting topic in the field of synthetic biology. Photocaging is still a very new and innovative strategy to work with. So it is possible to break down the topic of synthetic biology into a problematic aspect of our daily life. Do the test: can you explain the project to your family in a few sentences? I think you can do this.

Please name tree terms that describe the iGEM teams in Aachen and their work?
Self determination
Stick to details
Application orientated work

Could You please tell us something about the previous work of the iGEM teams in Aachen?
The success of the iGEM team 2014 was very unexpected and a great experience for all involved. This success motivated many and raised the attention for the iGEM team Aachen. The iGEM team 2015 repeated the success in a very similar way, showing how ambitious the members of the iGEM team Aachen are.

How do You feel to be a part of the iGEM team Aachen 2016?
Great! When the chance evolved to start an iGEM team in Aachen I was directly very positive. The interests of students and the idea of teaching are combined in the iGEM competition. Also I am very happy to see how the iGEM competition developed in the past few years, and I am excited how it will continue. I think it is also good that many students participate with fun and passion. In total iGEM is a great experience for me.

Can You tell us something about the atmosphere in Boston and at the Giant Jamboree?
The atmosphere is unique. All the teams live in the same hotel and when you enter into the convention centre, you can feel the iGEM spirit immediately. You will be very focused before you present your project in front of people, but afterwards you will just feel relieved and can enjoy the atmosphere. Besides it is great to represent the RWTH Aachen University and spend time with your team members. It is important to be recognized as iGEM team Aachen till the competition is over. Moreover, it is great to feel the international spirit. You can learn about so many projects and you have the chance to talk to many other teams about their iGEM experiences. Do not only look at the well-known teams, but talk also to teams with very difficult working conditions. Their enthusiasm is truly inspiring. In summary, enjoy Boston!

Toxicological Assessment

In order to evaluate the safety of our LIPs, we made a toxicological assessment, considering how humans and the environment could be affected when getting in touch with it and how probable an exposure is. We collected all data we could on the substance we want to replace. Furthermore, we examined the cleavage product of the protection group we intended to use. We compared the required amounts of chemicals with the existing solution to the amount that would be needed for ours as well as the dangers those substances pose.

Exposure

In our daily lives we get in touch with the contents of liquid washing detergents in several situations:

	A. Production During production of washing detergents, the factory workers are potentially exposed to high doses of the inhibited protease. There is a risk of dermal contact and inhaling the compounds.
	B. Home usage When washing at home the washing detergent can be spilled onto the skin of user. Thereafter residues of the washing liquid can remain in the clothes because of incomplete rinsing and cause dermal exposure.
	C. Drinking water Waterborne substances like boric acid, which cannot be filtered out in purification plants expose consumers via their drinking water.

Toxicology

Toxicology of boric acid
dermal:
LD₅₀ rabbit : > 2,000 mg kg [1]

oral:
LD₅₀ rat : 2,660 mg kg [2]

Boric acid reacts with the polyhydroxyl ribitol side chain of riboflavin. This increases its water solubility and reduces the amount of available riboflavin in animals and humans [3]

fatal dose human: [4]

2,000 - 3,000 mg for infants
5,000 - 6,000 mg for children
15,000 - 20,000 mg for adults

inhaling:
LC₅₀ rat : > 0.16 mg L ∙ hr [5]

reprotox:
Boric acid is classified as substance of very high concern [6] and is toxic for fertility. [7-8]

Toxicology of photocaged amino acids and their cleavage products
ONBY (Ortho-nitrobenzyl-tyrosine) and DMNBS (dimethyl-nitrobenzyl-serine) and their cleavage products ONB (2-nitrosobenzaldelyde) and DMNB (Dimethyl-nitrosobenzaldehyde) are not yet characterized in toxicology. Approximate predictions can be made through known data about similar chemicals as other nitroso compounds.

Figure 1: ONB-tyrosine cleavage reaction

Figure 2: DMNB-serine cleavage reaction

Nitrosobenzene
Nitrosobenzene reacts in the human blood circulation with hemoglobin and minimizes its ability of O₂ uptake.[9] Acute and chronic toxicity data are unavailable.

Other protection groups
To avoid the risks of the nitrosobenzyl derivatives there are a lot of different photo protection groups, which could be used instead of the ONB and DMNB.[10]

Comparative toxicological risk assessment

In absence of solid toxicity data predominantly for the cleavage products of the photo protection group of the enzyme it is difficult to impossible to conclude on the comparative risk assessment.

Assuming that both boric acid and the photo cleavage degradation product have similar toxicities the project approach has a clear advantage because of the much lower concentration in the product. While there are 16 mmol boric acid per kg laundry detergent the concentration of the photocaged amino acid is only 3,6 nmoles per kg. For every photocaged molecule of amino acid one molecule of protection group exits.

The common liquid laundry detergent contains 0.5 - 1% (w/w) of boric acid and 0.00005 - 0.0001% (w/w) active protease. [11]

Molecules of subtilisin in 100 g of liquid laundry detergent
For the molecular weight of subtilisin E we pasted our known amino acid sequence of 275 AAs into a calculation tool, the DNA sequence can be seen in part BBa_K2020023 [12]

weight of 1 molecule subtilisin E = 27.02 kDa

M (subtilisin E) = 27701.06 g mol
m (subtilisin E) = 0.0001% ∙ 100 g = 0.0001 g

n (subtilisin E) = 0.0001 g 27701.06 g mol = 3.6 ∙ 10^-9 mol

molecules of boric acid in 100 g liquid laundry detergent

M (boric acid) = 61.83 g mol
m (boric acid) = 1% ∙ 100 g = 1 g

n (boric acid) = 1 g 61.83 g mol = 0.01617 mol

molecules of boric acid per molecule of subtilisin E

n (boric acid) n (subtilisin E) = 448300

Conclusion

As the concentration of cleaved photo protection groups is almost 480,000 times lower than the concentration of boric acid its toxicity is allowed to be 448,000 fold higher for the same toxicity of the liquid washing detergent. If the toxicity of cleavage products is less than 448,000 fold higher than the toxicity of boric acid, washing detergents with replaced boric acid were less toxic than the actual.

Example for one washing cycle (100 g liquid laundry detergent)
For this calculation we assume that 1% of the liquid detergent stays in the laundry after washing and the composition is like in the calculation above.

DMNB and ONB group
n (cleaved group) = n (subtilisin E) = 1% ∙ 0.0001 g 27701.06 g mol = 3.6 ∙ 10^-11 mol
M (DMNB) = 195 g mol
M (ONB) = 135 g mol
m (DMNB) = 7 ∙ 10^-9 g

m (ONB) = 4.9 ∙ 10^-9 g

Boric acid
1% ∙ 1 g = 0.01 g

In this scenario 0.1 g of boric acid or 7 ng of DMNB or 4.9 ng of ONB remain in the laundry.

Comparative environmental risk assessment

The diluted compounds of washing detergents end up in the waste water and consequently in the waste water treatment plant.

Boric acid is highly water soluble and can be neither eliminated in the waste water treatment plant nor in the drinking water purification plant. [13]

The LIPs products are organic products, which could be eliminated by a biological waste water treatment plant but this remains to be verified as there is no related data available. As organic compounds they could most likely be filtered by active charcoal and others whereby human exposure by the drinking water can be excluded. Also, organic compounds will likely be degraded over time.

Laboratory Satefy Aspects

Used organism strains

Escherichia coli DH5α
Escherichia coli BL21 DE3
Saccharomyces cerevisiae
Bacillus subtilis

Potential risks

Our project poses the typical risks of working in a biology lab. So we reduce those risks by fulfilling safety level 1 procedures as seen below.

Waste treatment
All biological materials (including genetically modified organism (abbreviation GMO)) or equipment, that was used for handling, is collected separately and autoclaved.

Transportation:
Closed boxes are used for transportation between labs but if possible, transportation between labs should be avoided.

Emergency reaction:

If lab coats or clothes start burning, use emergency showers.
If acids or other harmful liquids get into the eyes, use eye showers.
If injuries occurred, treat them with first aid-kits, report the injury, and go to the hospital if necessary.
If solutions with GMO is running down the bench (or other kinds of contamination), swap it and disinfect the place with Bacillol.

Protective equipment:

Lab coats are mandatory.
Safety glasses are mandatory in special areas of the lab and should be used in other areas if necessary.
Usage of safety gloves are mandatory in special areas of the lab and should be used depending on the working materials.

Other rules:

Do not work alone. There should be at least one person from the lab, who is not part of the iGEM team.
Do not pipette with your mouth.
Disinfect hands after contamination with GMOs.
Disinfect hands and wash them after working in the lab.

Furthermore our project includes the handling of non-canonical amino acids, which we treat according to their special requirements.

Laws and regulations regarding biosafety in germany

References

[1]R. Krieger, “Handbook of Pesticide Toxicology”, Academic Press, San Diego California, volume 2, no. 2, p. 1414, 2001. [2]R. J. Sr. Lewis, “Sax's Dangerous Properties of Industrial Materials”, Wiley-Interscience, 11th Edition, p. 536, 2004. [3]J. T. Pinto, R. S. Rivlin, “Drug-Nutrient Interactions”, vol. 5, pp. 143-51, 1987. [4]R. Krieger, “Handbook of Pesticide Toxicology”, Academic Press, San Diego California, volume 2, no. 2, p. 1430, 2001. [5]European Chemicals Bureau, “IUCLID Dataset for Boric Acid”, 2000 CD-ROM edition, p.26, Oct. 2011. [6]European Chemicals Agency, “Comments and Response to Comments on Annex XV SVHC: Proposal and Justification”, 2010. [Online]. Available: https://echa.europa.eu. [Accessed: 19-Oct-2016]. [7]Human Health and Environment Task Force, “Human and Environmental Risk Assessment on ingredients of Household Cleaning Products”, 2015. [Online]. Available: http://www.heraproject.com. [Accessed: 19-Oct-2016]. [8]R. E. Chapin and W. W. Ku, “The reproductive toxicity of boric acid.,” Environ. Health Perspect., vol. 102, no. Suppl 7, pp. 87–91, Nov. 1994. [9]H. U. Käfferlein et al., “Bildung von Methämoglobin durch Anilin”, IPA-Journal, pp.26, Jan. 2014. [10]P. Klán et al., “Photoremovable Protecting Groups in Chemistry and Biology: Reaction Mechanisms and Efficacy,” Chem. Rev., vol. 113, no. 1, pp. 119–191, Jan. 2013. [11]Marktforschung Dalli-Werke Stolberg GmbH & Co. KG [12]Science Gateway, “Tools: Protein Molecular Weight Calculator”, [Online]. Available: http://www.sciencegateway.org. [Accessed: 19-Oct-2016]. [13]Barufke et al., “Bor: Ableitung einer Geringfügigkeitsschwelle zur Beurteilung von Grundwasserverunreinigungen“, LUBW. Landesanstalt für Umwelt, Messungen und Naturschutz Baden-Württemberg, Feb. 2012

Team:Aachen/HP/Silver