We were concentrating on the problematic substances diclofenac (analgetic) and azithromycin (antibiotic). Both substances pose environmental hazards in high concentrations. They reach our wastewater in large quantities. At wastewater treatment plants, only a small part can be degraded endangering aquatic fauna and flora. Other substances, such as ibuprofen or estrogen are also problematic. We utilize enzymes that can modify such substances rendering them less toxic. We chose the two laccases CueO (E. coli ) and CotA ( B. Subtilis ) for diclofenac, as well as the esterase EreB ( E. coli ) for antibiotics like erythromycin and azithromycin. Laccases are oxidoreductases that can oxidize phenolic compounds like in diclofenac, as well as other pharmaceuticals, such as carbamazepine, 17β-Estradiol. The resulting products are demonstrably less toxic. EreB has already been shown to being capable of breaking down erythromycin effectively. EreB also displays some promiscuity towards azithromycin. To increase the activity of EreB towards azithromycin we want to utilize site-directed mutagenesis. With the selected enzymes we are already able to degrade a wide range of pharmaceuticals. Besides the already mentioned substances, chloramphenicol, bisphenol A and 4-nonylphenol can also be oxidized by laccase. Enabling us to adapt our biofilm to different local conditions of wastewater pollution. Because of this B-TOX is not restricted to one area.

Where is the biofilm? Why is the Biofilm? How is the Biofilm

How are you going to solve the problem? Where did the idea come from?

Use this section to explain whatever you would like! Suggestions: Just Contributions.
[O.K.]

• Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua.
• Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua.
• Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua.
• Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua.

Achievements: (medal criteria)

Bronze

Competition Deliverables: We registered for the virtual Giant Jamboree and are looking forward to the event! Therefore, we created our Wiki, Poster, Presentation Video, Project Promotion Video and filled in the Judging Form.
Attributions: Please visit our Acknowledgements to see who supported us this year.
Project Description: We present ourproject description on our Wiki and a short version can be seen at “Introduction”.
Contribution: We contributed a variety of things to the iGEM community this year. Click on “Contributions” to see.


Silver

Engineering Success: We carefully designed our parts and show how to test them and evaluate results.
Collaboration: We collaborated with different teams throughout the year, like thepostcard collaboration with Duesseldorf, iJET with Aachen, Oxiteers with Stuttgart and Kaiserslautern and many more.
Human Practices: We engaged with experts and the public during the course of our project. We were able to generate a lot of valuable input and insights.
Proposed Implementation: We explain how our project will be implemented and give an outlook on alternative implementations, e.g. aboard the ISS.


Gold

Integrated Human Practices: We spoke to different groups of interests and ages, addressing their worries and incorporating their ideas in our project design. Especially, regarding the biosafety aspect of our project.
Project Modeling: We modelled our enzymes and fusion enzymes, biofilm growth and formation, and our kill switch. Together with iGEM Hannover we developed a software tool to model biofilm formation.
Partnership: We engaged in a partnership called the Oxiteers, with iGEM Stuttgart and iGEM Kaiserslautern.
Science Communication: We developed several strategies to educate society and make our efforts accessible for everyone.


Awards:

Best Education

We started a publicly accessible survey covering the opinion and knowledge about SynBio within Germany. Based on this survey, we broadcasted a livestream with iGEM Kaiserslautern to enable an open dialogue and interact with people all over Germany. We provided information to people who sought more knowledge and talked to them about their concerns regarding water pollution.
In our German podcast “Genomenal - Ein Haufen Zellen redet über Biotech” (English: Genomenal – a bunch of cells talk about biotech), we address ethical and legal questions in biotechnology. We provided our listeners with ways to easily communicate their thoughts to us and included their feedback. Additionally, we created a “how to podcast” guideline for anyone with the same intention covering necessities like equipment needs and how to build up your own RSS-feed.
We programmed a jump-and-run computer game called “The Genomal Adventures of Dr. W” together with Aleksa Zečević to playfully spark interest in biology even among the youngest of our audience.
The renowned German scientific journal BIOspektrum published our article, enabling us to reach an audience of 15,000 monthly readers. The BIOspektrum is linked to many German Societies in science such as GBM, VAAM, GfG or DPGT. As a result of this publication, Prof. Dr. Möller, astromicrobiologist at the German Aerospace Center, reached out to us to talk about possible implementation scenarios of our biofilm in aerospace and the adjustments necessary to enable its usage e.g. aboard the international space station (ISS).


Best Integrated Human Practices

We reached out to experts like Dr. Ulrich Ehlers from the Federal Office for Consumer Protection and Food Safety who also suggested the use of a kill switch to enable a safer use of our project.
As Prof. Dr. Sibylle Gaisser from the department of industrial biotechnology in Ansbach recommended, we created a safety form with the goal to prevent misuse of our biofilm, as well as a safety sheet to educate employees of wastewater treatment plants on biosafety issues.
We talked to many experts about all aspects of our project and used the information they provided us with to make our project come to maturation.


Best Model

We developed a software to model biofilm growth in collaboration with iGEM Hannover. It incorporates e.g. growth rates, split length or movement speeds of bacteria, enabling us to create plots of their position and movement as well as the robustness of the biofilm. It is based on literature values although experimental data can be easily integrated.
We used MATLAB to model our quorum sensing-based kill switch mechanism using ordinary differential equations (ODE) to make predictions about its functionality. It is also based on literature values with the intention to incorporate experimental data as soon as possible.
We determined possible 3D structures for one of our degradation enzymes EreB using RosettaCM. Using GROMACS for molecular dynamics simulation (MD), we validated the stability of our enzyme structure. With RosettaCM, we also predicted the structure of our fusion proteins of TasA and one of our degradation enzymes and simulated their binding affinity to their targets.


Best Software

We developed a software tool in collaboration with the iGEM Team Hannover which enables the user to run a molecular dynamics simulation of biofilm growth. It includes a three class and utility function. One of which is the “bacteria” class which is an object-orientated representation of the bacteria in the biofilm. Secondly, there is the biofilm class, which simulates interactions forces using a biophysical potential, and drag force and finally the “constants” class, which represents the interface the user utilizes to specify simulation constants like duration, step size or output paths. Other constants like initial population size and bacterial strain (Escherichia coli or Bacillus subtilis) may also be chosen by the user while the addition of one’s own parameters allows for easy simulation of one's bacterial population of interest. The code is open for everyone to use and our Wiki provides a detailed guide.


Best Sustainable Development Impact

The focus of our project to reduce wastewater pollution by pharmaceuticals is in strong agreement with the United Nation’s sustainable development goals 6 and 14. To ensure access to water and sanitation for all, especially, as in target 6.3, to improve water quality by reducing pollution and minimizing the release of hazardous chemicals is part of goal 6 and an important part of our project. We designed this biofilm for the purpose of wastewater treatment without complex and costly technological adjustments but with the versatility of bacterial biofilms. We can see the consequences of releasing pharmaceutical pollutants into the environment as it has been reported to endanger several species in the recent past. That is why goal 14 inspires us to conserve and sustainable use the oceans, seas and marine resources and to significantly reduce marine pollution as covered in target 14.1. It is of great importance to us to prevent harm to our marine ecosystems due to contamination of the environment with pharmaceutical pollutants.

Use this section to explain whatever you would like! Suggestions: Outlook