- Register for iGEM, have a great summer, and attend the Giant Jamboree
We registered and was accepted by iGEM Headquarters on 2016-05-10 at 09:08:56. We definitely had a great summer, and we are excited to attend the Giant Jamboree.
- Meet all deliverables on the Requirements page
We sure did. You are reading our wiki right now. All our parts were registered and submitted. The safety, judging and registry forms were filled as required. Also, a poster is ready and the team members are set to present our work at the Giant Jamboree.
- Clearly attribute each aspect of the project
All the help that was needed is greeted with great gratitude under Attributions.
- Document at least one new standard BioBrick Part or Device central to your project and submit this part to the iGEM Registry.
K2018030 is a secretion system for poly-beta-hydroxybutyrate (PHB). This secretion tags PHB with a hybrid protein consisting of phasin, that binds PHB and a hemolysin tag, and is recognised by a type 1 secretion system. The BioBrick also contains two transport proteins that will recognize the hybrid protein and secrete it along with PHB. This construct creates the possibility for a large scale production of PHB in a continuous stirred-tank reactor.
- Experimentally validate that at least one new BioBrick Part or Device of your own design and construction works as expected
Our new devices K2018014 and K2018015 are hybrid bacteriocins that show great effect at inhibiting growth of multi-resistant Methicillin Resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa. The hybrid bacteriocins show greater effect than the native bacteriocins.
We have helped the following iGEM teams in different ways: The iGEM team from the Technical University of Denmark, the University of Copenhagen, the Chalmers University of Technology, Stockholm, Virginia and LMU TUM Münich. For more details, please see Collaborations.
- Human practice
As it was mentioned by iGEM, this competition is much more than just sitting on the lab bench. We have not only been reaching out to highschool students at different events, but we also managed to create videos to educate the Danish citizens about synthetic biology. Futhermore, we considered bioethics and philosophy of science in the methods of our project. You can read more about it under Practices and Prospects.
- Integrated human practice; Expand on your silver medal activity of human practice by demonstrating how you have integrated the investigated issues into the design and/or execution of your project
We talked to several experts in order to develop new goals for the future perspective of our project and to expand our knowledge about the different sub elements. Among those are experts in the areas of therapeutic use of biodegradable plastic and experts in the use of antibiotics and bacteriocins as new antimicrobial components. Furthermore, we made a survey to one of the high schools we visited and used their feedback to improve our presentation. They also gave us new ideas for future development of our Bacto-Aid. Moreover, we talked to a nurse about their requirements for a patch like ours. Lastly, we made a survey to Danish companies that work with plastic production and investigated the possible interest in the use of plastic produced by bacteria. For more information go to Integrated Human Practice or explore our website to see where it was integrated.
- Improve the function OR characterization of an existing BioBrick Part or Device and enter this information in the Registry
We examined the phaCAB BioBrick created by Imperial College team 2013. We characterized the relationship between the strength of the additional promoter and ribosomal binding site and the yield of plastic. In the process we created another phaCAB construct that appears to generate a higher yield of PHB. Furthermore, we characterized the effect of the pantothenate kinase II BioBrick submitted by Imperial College’s 2015 iGEM team on E. coli’s proteome. For more information see our Demonstration & Results Page.
- Demonstrate a functional proof of concept of your project. Your proof of concept must consist of a BioBrick device; a single BioBrick part cannot constitute a proof of concept
By producing bacteriocins and PHB from different BioBrick devices, we demonstrated a functional Proof of Concept. Furthermore we have a BioBrick device (K2018050) that secretes PHB.
- Show your project working under real-world conditions. To achieve this criterion, you should demonstrate your whole system, or a functional proof of concept working under simulated conditions in the lab
Through MIC tests of our bacteriocins, we showed their effect towards different human pathogens that are particularly complicated to treat due to the rising resistance problem. These tests suggested the potential of these bacteriocins to be used in therapeutic practice. We have improved the production platform of PHB by introducing a secretion system for PHB in E. coli. Using PHB produced with our construct, we 3D printed an actual product; an implant prototype of the human mandible (jaw). This illustrated that PHB produced in bacteria can be used for real world applications. To read more about this, go to Demonstration & Results and Proof of Concept.
- Education and public engagement
During the project, one of our main focuses was to engage ourselves in the community and in discussions with whomever interested in synthetic biology. We held several talks and presentations. The audiences varied from small children at the Research day at the University of Southern Denmark to professors in philosophy. We taught a lot and learned even more ourselves. One of our engagements was being teachers for a day: we educated two biotechnology classes at a local high school an entire day. We taught them about synthetic biology and made a laboratory exercise for them, which we supervised for. It was great fun. We also focused on taking a stand for scientific reproduction, which we did further research about. The biotechnology classes tried to reproduce some of our protocols, which were mostly successful.
- Integrated Human practice
We talked to several experts in order to integrate new goals for the future perspective of our project and to expand our knowledge about the different sub elements. Among those are experts in the areas of therapeutic use of biodegradable plastic and experts in the use of antibiotics and bacteriocins as new antimicrobial components. We tried to integrate some of their ideas into our project in order to make our Bacto-Aid more realistic to commercialize. The interviews also reassured us, that we are dealing with a very essential problem. Furthermore, talking to the experts about PHB, we found a great interest regarding PHB as therapeutics, which we were unaware of. This expanded the potential uses of PHB in our project. This lead us to 3D print a life-size part of a jaw.
We talked to a nurse about their requirements for a patch like ours. This guided us regarding to which aspects we should focus on if hospitals are going to use our Bacto-Aid. Furthermore, we made a survey with one of the high schools we visited and used their feedback to improve the message. Lastly, we send out question forms to Danish plastic companies to investigate their possible interest in the use of plastic made by bacteria. They were very interested in our plastic, but today PHB is too expensive. We have therefore worked hard to optimize the production of PHB.
- Best device
We created a BioBrick (K2018050) that will help overcome the problems associated with extraction and production of PHB. The Biobrick allows the PHB producing E. coli to secrete internal PHB into the environment. The BioBrick utilizes the type 1 secretion pathway for hemolysin and tags PHB for secretion by fusing a part of hemolysin to the PHB-binding protein, phasin. The BioBrick has been shown to work in Top10/PanK-sec. The PHB secreting cells could enable optimal utilization of a continuous system for large scale PHB production.