Before we started planning the tasks for each sector of our project, we spent a lot of time brainstorming the goals behind the activities within them. We thought about what we should do to make our plan responsible and good for the world.
At first, the Human Practices sector seemed to us the most incomprehensible. However, we eventually realized that Human Practices endeavours cannot stand alone. Human Practices are a complex representation of the world experiencing the problem we want to solve.
An analogy for a representative form of this sector could be a mechanical watch. Human Practices is its dial, an element that enables the display of the hour, which results from the watch mechanism's complex work. In the analogy presented, the displayed hour is the goal of the project: a way of solving the problem we have undertaken. This goal can be successively achieved thanks to the work of the other sectors - lab, education, and social media - which are the cogwheels, on whose mutual relationship depends the final effect of the actions we have undertaken.
Only some people are watchmakers, so not everyone needs to understand the mechanism of a watch. However, time, the effect, concern all of us. This is why the actions taken by Human Practices are so crucial to us. Things we do have a real influence on shaping the world, so our priority is to live up to its expectations.
The city of Gdańsk, where the majority of our team members live, is located in the northern part of Poland on the Baltic Sea. Due to our location, we spend a significant part of our free time on seaside walks, and therefore we have direct and tangible contact with marine water pollution. We often saw trash breaking on the shore, littering the beach, or making it difficult for coastal birds to move.
The proximity of the Baltic Sea inspired us to focus on local problems of marine water pollution and their impact on human and aquatic health and quality of life. The trash we saw on the beach was a visible, tangible problem, but because of our field of study, we are used to noticing less visible, potential problems that can exist at a micro-scale, thus revealing their real harmfulness. The presence of trash on the seashore undoubtedly affects the cleanliness and safety of the water, so we decided together that the problem we would like to address as part of our competition project would be related to the neutralization of water pollution.
Initially, our interest was focused on the problem of water pollution with microplastics. However, during our literature review and research, we came across two projects that are being implemented locally in Gdańsk: NonHazCity and NonHazCity2. The main goal of the projects is to educate and create innovative solutions to minimize the emission of hazardous substances from urban areas to the Baltic Sea. As part of the project, wastewater and rainwater were examined for the presence of compounds that are toxic to living organisms. The focus was primarily on substances that affect hormonal balance - endocrine-disrupting compounds. [1]
In this way, we encountered a problem with which we had no previous direct contact - environmental pollution with phthalate derivatives. This inspired us to take action and address a problem that is not as widely reported in the media as microplastic pollution.
Therefore, we decided to design a filter that contains immobilized enzymes that degrade phthalate derivatives to safe, commonly used in the food industry, benzoic acid.
We brainstormed and conducted research to learn how the presence of endocrine-disrupting compounds in the environment affects the health of animals, including humans.
We learned that phthalates, as endocrine-disrupting compounds, have a negative impact on the endocrine and reproductive systems, contributing to the development of diseases associated with hormonal imbalances, as well as increasing the risk of infertility. [2,3] We consulted the information we obtained with Dr. Dominik Rachoń, an endocrinologist from the Medical University of Gdańsk, who told us about the impact of EDCs, including phthalates, on the health of the endocrine system. More details on this topic can be found in the subsection:
Phthalates are used in industry as plasticizers - they improve the mechanical properties of polymers used to produce bottles, increasing their flexibility and bending ability, which is why bottled water is one of the main sources of human exposure to phthalates. Phthalate derivatives do not form strong bonds with polymers, so they easily enter the aquatic environment. In addition, phthalates serve as solvents and emulsifiers, and they can be found in kitchen utensils, medical devices, toys, cosmetics, clothing, building materials, electronic devices, and dust. [4,5]
According to Directive 2013/39/EU of the European Parliament and of the Council, DEHP is classified as a priority hazardous substance, but no maximum permissible concentration (MPC) for this substance has been set for surface waters. According to the United States Environmental Protection Agency (EPA) guidelines, the MPC for DEHP in drinking water is 6 μg/L. [4]
The World Health Organization (WHO) has classified DEHP as a potentially carcinogenic substance (group 2B) with a safe MPC for humans of 8 μg/L. [2]
The highest concentrations of DBP and DEHP in bottled waters from 21 countries were 222.0 mg/L and 94.1 mg/L, respectively, meaning that the DEHP concentration was almost 12,000 times higher than WHO standards allow. [6]
In 2020, the European Union Directive 2020/2184 established reference values for drinking water quality, but none of these limits apply to phthalate derivatives. [4]
Conclusion: The threat posed by the presence of phthalates in the environment is global, so we believe that it is a social responsibility to create a tool that will enable the elimination of phthalates from the environment.
We are aware that pollution of the environment with phthalates, which belong to the group of endocrine-disrupting compounds, is only a drop in the ocean of problems that the contemporary world is already facing or will face. However, we believe that a comprehensive approach, focused on solving specific problems, is an important step towards achieving all 17 Sustainable Development Goals.
Our filter project fits into four of the 17 Sustainable Development Goals [SDGs] campaign:
As we have already mentioned, the problem of environmental pollution with phthalates is a global one. Water is a key component of the world, without which it could not exist in the form we know it. Water is needed by all organisms, for some it is a living environment, and it is the basis of all industries, such as food or pharmaceutical. Phthalates, which are not eliminated from the aquatic environment, can lead to hormonal disorders and infertility among all users of water, which is obviously a cause for concern.
The use of water concerns every aspect of life, therefore the need to care for its purity and reduce the damage caused by its pollution has a global character, and the benefits of their elimination concern all its users.
We believe that our filter can be useful on two levels, which differ in scale:
We assumed that the use of our filter at the industrial level - as an element of the infrastructure used to clean water from pollutants - would make it possible to limit the circulation of phthalates in nature. Water free of phthalates would reach natural water bodies, which would make it possible to protect aquatic flora and fauna. The use of the filter in wastewater treatment plants would also make it possible to provide safer, better quality water to all industrial sectors and households.
To convince ourselves that our assumptions are correct and to find out what the demand for a similar product is, we visited a local wastewater treatment plant in Toruń. [link]
In addition to the visit to the treatment plant, we consulted with representatives of Bioton, who gave us valuable advice on the scalability of the product and its safety. More on this can be found in the entrepreneurship section.
We believe that our filter could be used as a consumer product - in the form of a water filter bottle or filter for bottles, providing access to clean, phthalate-free water to each of its users. Thanks to this potential application, everyone could take care of the safety of the water they drink on their own, minimizing the risk of exposure to harmful substances.
To find out what the social attitude towards our project is, we took part in events such as the Open Day of the University of Gdańsk or the Fahrenheit Picnic, where we had our stands, allowing us to interact with people who could potentially be users of the filter. Public engagement activities, which are described in more detail in the Education section, allowed us to have many valuable conversations and learn about social expectations for the product we want to create.
During the Open Days of the University of Gdańsk, which took place before the start of our planned work, we conducted a survey that allowed us to gather many valuable opinions that greatly influenced the design of our product.
In addition, as the local character in the context of the Baltic Sea does not only concern Poland, we conducted a survey about our project among teams from the Baltic countries.
One of the ways our team used to learn about the community whose problem we want to solve was to conduct surveys on our project, endocrine disrupting compounds, marine water pollution, and available methods for their elimination. The results of the surveys, which were feedback on our idea, gave us the opportunity to learn about the concerns and expectations of society regarding our filter, thanks to which at the design and planning stage of the experiments we could introduce changes that meet the real needs of our potential future recipients.
We believe that together with the people affected by the problem we want to solve, we can create something that will have a positive impact on the future of the world.
During the preparation of the questionnaires, we paid particular attention to legal regulations and ethical standards related to human subject research. Our surveys did not use sensitive data, so we did not have to have the approval of the Polish ethical committee to conduct them and to legally use the data obtained. Each of our surveys began with content informing the respondent what the purpose of the questionnaire is and how the data obtained will be used. We repeatedly consulted the designs of our surveys with people more experienced in conducting them, to make sure that they do not violate ethical norms, are transparent and understandable. Thanks to this, we learned, among other things, how important it is to include consent to data processing.
During the Open Days of the University of Gdańsk, we conducted a survey aimed at event participants - among them, high school students dominated, but it was not the only age group. After all, we received responses from 57 people in the 16-21 age range.
The results of the survey provided us with valuable information about society, concerning knowledge about the harmfulness of plastic packaging, water pollution, available methods for reducing these pollutants, and genetic engineering.
The results of the survey showed us that practically all respondents are aware of the level of pollution of surface waters - which convinced us of the need to develop a product that eliminates the problem of pollution. Additionally, the results gave us to understand that the majority of the local community has concerns related to drinking water from plastic bottles, having limited trust in its purity.
In addition, we learned that public awareness of environmental pollution by a group of endocrine disrupting compounds, including phthalates, is low - which allowed us to adapt educational activities to the real needs of society. More on this topic can be found in the Education section.
Most of our respondents expressed negative views about the impact of endocrine disrupting compounds on the body, signaling to us that our iGEM competition project to eliminate these compounds from the environment is addressing a real social need.
The survey was conducted at the first stage of planning the activities related to the actual implementation of our idea - it therefore had a huge impact on the final design of our filter. Before conducting it, our idea included the use of genetically modified bacteria capable of degrading phthalates, which would be placed in the filtering part of our device. After familiarizing ourselves with the results of the survey, we noticed that the consumption of water purified by a bacterial filter raises social concerns, in connection with which some of the survey recipients, the group affected by the problem we want to solve, do not express a desire to use a filter that contains bacteria. As our assumption is that the problem of environmental pollution by phthalates is global in nature and the filter we are designing should meet the expectations of as large a part of society as possible - the concerns we noticed prompted us to reflect and take appropriate actions that are a response to the presented expectations.
The information we gathered from the local community helped us to shape the focus and direction of our project. It also helped us to develop a product that is more likely to be successful and accepted by the people who need it most.
Social concerns about drinking water filtered through a bacterial filter led us to abandon the idea of a phthalate removal system using genetically modified microorganisms.
The main reason we decided to address the problem of water pollution by phthalates is that it largely affects the Baltic Sea, which is a local concern for us due to Poland's location. However, Poland is not the only country associated with the Baltic Sea - therefore, in our project we wanted to consider the opinion of residents of other Baltic countries that face the problem we want to solve.
In this regard, our team prepared a survey addressed to teams participating in the iGEM competition coming from countries neighboring the Baltic Sea. In the survey, we wanted to examine awareness about water pollution and its effects - which allowed us to adapt educational materials to the level of knowledge of our audience. Above all, we wanted to survey opinions on how we want to solve the problem of pollution in EDCs - what is the attitude of other communities, other than our local one, to a product that should meet the expectations of society on a global scale.
We received responses from 14 participants from four countries bordering the Baltic Sea: Germany, Finland, Latvia and Sweden. Unfortunately, we were unable to obtain responses from teams from Denmark, Estonia and Lithuania.
All of our respondents were aware of the pollution of surface waters, and most of them are also aware of the level of pollution of our local water body, the Baltic Sea.
Among the local causes of the increasing level of pollution of the Baltic Sea waters, our respondents mentioned, among others: industrial pollution, chemical fertilizers, pesticides and herbicides. Members of teams from Sweden and Lithuania confirmed the presence of water quality protection programs in their countries, the remaining respondents were unable to comment.
All of our respondents were aware of the pollution of waters with microplastics, however, most of them had not yet encountered the term endocrine disruptor pollution, including the problem of pollution of the environment by phthalates. Nevertheless, the negative attitude and concerns expressed towards these terms gave us a signal of the importance of the problem we want to solve. At the same time, the feedback we received allowed us to adapt educational content - especially in our social media - to the level of knowledge of society.
Part of the survey we conducted concerned our filter directly. The vast majority of respondents expressed positive opinions about the possibility of drinking water through a bottle containing a filter to which degrading enzymes are immobilized. We obtained valuable information about the expectations for our product, which included, first of all: safety guarantee, reduce exposure to endocrinally active water compounds, use of eco-friendly materials, ease of use.
Thanks to the survey, we also learned what concerns about our filter project have its potential future users. Valuable feedback will help us to choose the direction regarding the steps and limitations that we should overcome in order to improve our product so that we can implement it in the real world.
The feedback we received prompted us to reflect on what we should do, what changes to make in the further stages of design, in order to make our project safer and better for the world. Issues that we will have to consider before trying to bring our filter to market include:
Many thanks to the teams that participated in the survey - getting to know the problems of our coastal neighbours was very informative and contributed to many eye-opening reflections.
Special thanks to: Bochum iGEM Team, Stockholm iGEM Team, Aalto-Helsinki iGEM Team, ABOA iGEM Team, Latvia-Riga iGEM Team, Lund iGEM Team, Freiburg iGEM Team, Dresden iGEM Team, Kaiserslautern iGEM Team
Due to the global scale of water pollution by endocrine-disrupting compounds, we want our filter to be used in a way that protects the largest possible group of people. Therefore, we assumed that it could be used in wastewater treatment plants - places that ensure the quality and purity of water supplied to all people with access to sewers. To see if our assumption is justified in the real world and whether the filter we are designing could help to improve the safety of tap water, we decided to visit a local wastewater treatment plant in Toruń. We wanted to learn about the commonly used methods of water treatment, their effectiveness, and the limitations associated with the process. Most importantly, we wanted to know if there is a real need for innovations that reduce the problem of water pollution by endocrine-disrupting compounds (EDCs) - and if so, what conditions our filter would need to meet in order to be used in a wastewater treatment plant.
During our visit, we had the opportunity to see the entire wastewater treatment system in operation. We traced the entire process from the point where the contaminated water enters the plant to the point where the treated water flows out to the city's residents. We learned that wastewater treatment plants are not legally required to monitor the levels of EDCs in the treated water, and as a result, no methods are used to remove contaminants from this group of compounds. Heavy metals, whose concentrations are legally monitored, are primarily deposited in excess sludge, as are phthalates. Excess sludge is transported to thickeners and then to anaerobic digesters, where biogas is produced. In this process, pathogens and parasites are also killed. The resulting sludge is then used for composting or is converted into granules for soil fertilization. The form of its reuse depends on the final concentration of heavy metals - the Ministry of Environment Regulation sets standards for the concentrations of heavy metals that may be present in sludge, and the values of these standards determine the choice of location where it can be reused.
We learned that our filter could be used in the final stage of the wastewater treatment process, in the form of filter beds at secondary clarifiers. Due to the risk of contamination of the filter with sludge and waste, it would need to be properly protected from external environmental factors, otherwise its operation could be blocked.
Additionally, since phthalates are not among the hazardous substances that must be removed from water and whose concentration must be monitored on a regular basis according to the Wastewater Directive, wastewater treatment plants would not decide to use the filter until the current legal situation changes.
We were advised to direct our filter proposal to industrial enterprises that produce materials that use phthalates, such as plastic processing plants. Due to the release of hazardous substances into the environment, industrial plants are required to monitor the concentration of substances not covered by the Wastewater Directive. The information we obtained created a new perspective for the application of our filter and increased our awareness of the infrastructure in which it could be used, which will have an impact on the further stages of design.
This made us realize that it is necessary to implement activities to raise public awareness of the health risks posed by the presence of phthalates in water. The effectiveness in reducing this risk will require us to directly contact the relevant representatives of the legislative authorities to draw their attention to this problem and, in the long term, encourage them to introduce appropriate regulations that would require wastewater treatment plants to monitor the concentration of endocrine-disrupting compounds.
We learned that there is a particular need to raise public awareness about how to dispose of everyday items such as cotton swabs, kitchen paper towels, and wet wipes. Wastewater containing these products contributes to the clogging of screens separating solids from liquids in the first stage of mechanical wastewater treatment. This allowed us to create educational content that addresses the problems faced by wastewater treatment plants.
The analysis of the survey results conducted during the Open Days of the University of Gdańsk led us to reflect. First and foremost, we received feedback from the community regarding the use of microorganisms as a filter element responsible for degrading phthalate derivatives. The majority of the responses were positive about our original idea of creating a system to eliminate phthalates, but 8.8% of respondents expressed a negative attitude towards consuming water filtered through a bacterial filter. However, as we have mentioned many times before, the problem of environmental pollution with endocrine disrupting compounds is global. Therefore, the reluctance towards our proposed system for cleaning water from phthalate derivatives prompted us to take responsibility for creating a product that will be perceived as safe by the widest possible audience.
We believe that the best solution to this problem would be to create a system that does not contain living organisms, but only the enzymes that they would use to degrade phthalates. To this end, we decided to consult our ideas for switching to a cell-free system with an expert from our institute, Professor Michał Obuchowski.
During our conversation, we learned that the European Union has very strict regulations on GMOs. Cultivation and trade of GMOs are only permitted after a risk assessment that aims to ensure the safety of humans, animals, and the environment. After reviewing Directive 2009/128/EC on the prevention and management of risks associated with the use of genetically modified organisms in the environment and Directive 2001/18/EC on the placing on the market of genetically modified organisms, we concluded that the introduction of our filter into industrial use would be difficult to implement. Due to the legal situation of GMOs in the European Union and Poland, our idea could prove to be ineffective - legal restrictions would prevent its implementation in the real world, which would not reduce the problem of water pollution with phthalates.
Prof. Obuchowski discussed with us many potential ideas for creating a cell-free system that can effectively degrade phthalate derivatives present in water. We jointly concluded that the optimal solution would be to create a system using enzymes - an esterase and a decarboxylase, which would be immobilized on a platform that is a mini-cellulosome. The use of immobilized enzymes has allowed us to create a system with improved efficiency and increased stability - additionally, immobilizing enzymes on the platform is associated with a lower risk of releasing them into the environment. Thanks to the use of this system, the final product of phthalate degradation is benzoic acid, which is recognized as a substance safe for human health, which is why it has found application as a food preservative in the food industry. Enzymes intended for immobilization are produced in a system based on E. coli - however, thanks to the use of available methods for purifying enzymes produced in bacterial systems used in pharmaceutical companies, the final product is not legally perceived as GMO. More on this topic can be found in the BIOTON section.
WHAT WE LEARNED:
Thanks to the opportunity to consult with Prof. Obuchowski, we were able to create a new filter design, which allows us to eliminate the use of genetically modified bacteria - which contributed to the fact that the use of our filter is more scalable, as it does not fall under the legal restrictions imposed by the European Union. First and foremost, the new form of our filter meets the expectations of potential consumers, who expressed mixed feelings about drinking water purified by a bacterial filter. We care that our proposed product is perceived as useful and safe - the changes mentioned above have brought us closer to achieving the desired effect.
Due to the impact of phthalates, which belong to the endocrine-disrupting chemicals (EDCs), on the endocrine system, it was very important for us to speak to a healthcare professional who has a direct experience with the problem we are trying to solve. Professor Dominik Rachoń, MD, PhD, is a practicing endocrinologist and head of the Department of Clinical and Experimental Endocrinology at the Medical University of Gdańsk. He has devoted a significant part of his career to studying EDCs, particularly bisphenol A and its analogues, and their impact on the endocrine system. As a result, our conversation with him provided us with a solid foundation of knowledge and a realistic picture of the situation at the scientific and clinical level. Professor Rachoń has a broad knowledge of the molecular mechanism of action of EDCs, which helped us better understand why they are so dangerous to the human health. EDCs are structurally similar to steroid hormones, including estrogens, which play a significant role in reproduction and women’s health. As a result, they can interact with the receptors of these hormones, acting as agonists or antagonists, which can lead to several hormonal disorders. Through the discussion, we learned about the clinical picture of the selected endocrine disorders, the diseases most diagnosed in recent times, and the reasons for the global decline in fertility worldwide. Contrary to popular belief, endocrine disorders, such as thyroid diseases, are not currently the most common endocrinopathies. An important issue that Professor Rachoń drew our attention to, is the prevalence of self-diagnosis of thyroid diseases among patients, which results from the possibility of independently performing tests to check the levels of thyroid hormones and their precursors in diagnostic laboratories. This often leads to overinterpretation and erroneous, worrying conclusions. The most common endocrine disorder diagnosed in women is polycystic ovary syndrome (PCOS). Women with PCOS are characterized by an exaggerated ovarian androgen, which leads to several clinical consequences, including acne, hirsutism, and androgenic alopecia, but most importantly, to menstrual irregularities, which contribute to fertility problems. Professor Rachoń's scientific work includes numerous publications on the impact of exposure to bisphenol A and it’s analogues, on the development and clinical picture of PCOS. However, he advised caution in drawing final conclusions and emphasized the need for further research in this area. Professor Rachoń suspects that phthalates also belonging to the EDCs with affinity to steroid hormone receptors can also cause several endocrine and metabolic abnormalities. During the conversation, we also learned more about the decline in fertility in men and women worldwide, which may also be caused due to the exposure to phthalates. Professor Rachoń told us about the potential role of phthalates in this process, focusing on their mechanism of action. Phthalates stimulate estrogen receptors, which the pituitary gland interprets as a signal of estradiol production in the body. As a result, according to the feedback mechanism, the pituitary stops the release of the follicle-stimulating hormone (FSH), which is crucial for the development of the ovarian follicles in women and spermatogenesis in men. The laboratory reference values for the number of sperm in the ejaculate have decreased dramatically in recent years, which is indirectly attributed to the exposure to EDCs, including phthalates. Unfortunately, the study of the impact of EDCs on human health is difficult, due to the fact that these compounds can affect gene expression both in adulthood as well as in the prenatal period. Disorders that are the consequences of exposure to EDCs in the prenatal environment and early childhood may manifest themselves many years after intensified contact with compounds from this group, which limits the possibility of studying their impact on the development of various disorders during the lifetime. In addition, Professor Rachoń drew our attention to the fact that, in addition to synthetic compounds such as phthalates and bisphenol A, EDCs also include naturally occurring substances in plants, such as polyphenols and phytoestrogens, which can also affect the functioning of the endocrine system. He also pointed out that the number of synthetic compounds is constantly increasing, so the number of substances that may have endocrine-disrupting properties is progressively expanding. As a result, it is difficult to fully protect oneself from exposure to potentially dangerous substances. In life, we are exposed to a variety of different chemicals that affect the endocrine system, as well as to other physical factors i.e. UV radiation. It is difficult to say whether the development of a specific dysfunction is due to only one factor, or perhaps to a group of factors to which a given person is exposed.
WHAT WE LEARNED:
Thanks to Professor Rachoń's extensive knowledge of the molecular mechanism of action of EDCs, we were able to gain a deeper understanding of why they are so dangerous to the endocrine system. We believe that our project has the potential to make a significant impact on our society by helping to improve public health and reduce the burden of endocrine and metabolic diseases. We are grateful to Professor Rachoń for his insights and guidance, and we are committed to working to make our project a success.
Dr. George DiCenzo from the Department of Biology at Queen's University in Canada visited our university as part of the Visiting Professors program. Dr. DiCenzo's research focuses on genomics, metabolic pathways, and their regulation in symbiotic bacteria of the genus *Sinorhizobium meliloti*. His recent research also concerns an efficient approach to the biodegradation of plastics, which led us to consult him with our idea. During the meeting, we presented Dr. DiCenzo with our idea and our preliminary plans for its implementation. We received many valuable insights about the need to verify whether the end product of the degradation of phthalic acid compounds by enzymes is broken down by bacteria, which would make it more biodegradable.
WHAT WE LEARNED:
The conversation confirmed our choice of enzymes used in our filter - esterases and decarboxylases, which degrade phthalic acid derivatives to benzoic acid, which can be broken down by many groups of microorganisms, such as Pseudomonas, Syntrophus, and Streptomyces. (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7354641/)
Bioton is a Polish biotechnology company that provides comprehensive solutions for the treatment of diabetes. The company is involved in the production and distribution of biotechnology products - it is one of the 8 world producers (on a commercial scale) of recombinant human insulin and the undisputed leader in Poland. In addition, it produces generic drugs - mainly antibiotics. The consultation with Bioton enriched our team with knowledge about the process of introducing new technologies into the industrial production process - we received many valuable suggestions that would make our project more scalable and accessible to the industrial sector. During the meeting, we presented our project, current results, and ideas for further development. During the conversation, we also discussed the safety of conducting industrial biotechnology processes and the available methods of efficient purification of proteins produced in genetically modified organisms. More about the meeting can be read in the Entrepreneurship section. During the meeting, we received valuable tips on how to adapt our filter to the conditions of the industrial sector - which, due to the global nature of the problem of water pollution by phthalates, is particularly important to us.
WHAT WE LEARNED:
We learned that the possibility of using our filter on an industrial scale will require additional experiments, the results of which may affect the efficiency of the process of cleaning water from phthalates at an industrial level. Suggestions for further research that we would like to conduct include, among others, determining the stability of the bond between dokerin and cohesion in different environmental conditions - at varying pH and temperature. Bioton also proposed a solution to immobilize enzymes to by-products from wood processing plants - such as sawdust, which is mainly cellulose. Due to the biodegradability of this raw material, the use of this solution would allow for the simplification of the platform disposal system and the removal of used enzymes. We are excited about the possibility of implementing this system - it would make our idea even more environmentally friendly.
Dr. Katarzyna Węgrzyn is the PI of our team and a member of the Molecular Biology Department of the Biotechnology Institute at IFB UG & MUG. The research conducted in the Laboratory includes the analysis of molecular mechanisms responsible for DNA replication and the stable maintenance of extrachromosomal genetic elements. Experimental projects are carried out using in vivo experiments on living cultures of bacteria, in vitro in experimental systems reconstructed from purified elements, and in silico using bioinformatics techniques. In her research work, she uses cloning techniques using plasmid expression vectors - in the third semester of our undergraduate studies, she taught SnapGene software with Prof. Obuchowski.
What we learned:
Due to Dr. Węgrzyn's experience with the cloning procedure using bacterial expression plasmids, we consulted with her our ideas for the production of enzymes, which are the basis of our designed filter, in the E. coli bacterial system. We presented her with our problem related to restriction and ligation analyses, which appeared at the stage of planning experiments. We learned that the plasmids we originally selected are not compatible, which may contribute to the aforementioned complications - Dr. Węgrzyn suggested which expression vectors we should use to carry out efficient overproduction of the enzymes we want. Additionally, we learned that we should only determine the size of the plasmid in a linear form and the need to introduce a larger number of control trials. The help we received from Dr. Węgrzyn enabled us to design the experiments in a way that allowed us to clone the sequences of the enzymes we selected into the expression vector (which was selected?), thanks to which we were able to carry out their overproduction in the E. coli system.
Professor Stanisław Ołdziej is the head of the Biopolymer Structure Laboratory at the Institute of Biotechnology of the University of Gdańsk and the Medical University of Gdańsk. The research conducted in the Biopolymer Structure Laboratory focuses on the understanding and description of various types of intermolecular interactions that govern the very early stages of protein folding. In his work, he uses techniques such as mass spectrometry, NMR spectroscopy, and high-performance liquid chromatography. Professor Ołdziej helped us design HPLC experiments - he consulted our ideas, and pointed us to literature that allowed us to adapt this method to the requirements of our project. Our team received from him enormous substantive support, consisting of explaining the theoretical aspects of the HPLC method and providing advice on how to optimize our experimental method. Professor Ołdziej also provided us with technical support - he provided us with HPLC equipment and the necessary reagents, which allowed us to conduct experiments necessary to determine the function of the enzymes we produced.
Marcel Thiel, M.Sc., is an assistant at the Biopolymer Structure Laboratory of the Institute of Biotechnology IFB UG & MUG in Gdansk, Poland.
What we learned:
M.Sc. Marcel Thiel has experience in working with cleaning, purity testing, and protein identification using HPLC techniques. Therefore, he conducted a practical training session for us on how to use this method and helped us select the appropriate equipment before our research team began working with this procedure. Thanks to the fact that our work was carried out under the supervision of an expert, we were able to conduct our experiments safely and efficiently.
Dr. Dorota Krzyżanowska is a member of the Department of Biologically Active Compounds at the Institute of Biotechnology of the University of Gdańsk and the Medical University of Gdańsk, where she is the Lab Manager. The research conducted by the Laboratory covers topics in the field of production of biologically active compounds in plant in vitro cultures and the search and study of natural and synthetic antibacterial and antifungal compounds. In her work, she uses techniques such as classical microbiology, RT-qPCR and molecular cloning.
What we learned:
Dr. Krzyżanowska provided our team with technical and substantive support. All members of our team responsible for the experimental part of our project, attended a safety training in the laboratory, which was conducted by Dr. Krzyżanowska. In addition, we received support from her during the use of new chemical reagents necessary to carry out the experiments, which could pose a potential threat to us. Dr. Krzyżanowska explained to us where to find information about the risks and instructed us how to work with them - thanks to which we were able to carry out the project responsibly and safely. In addition, Dr. Krzyżanowska helped us fill out the Safety Forms application, so that our project can be legally conducted in Poland, taking into account all aspects related to its safety - both for team members and the outside world. Her support is described in more detail in the Entrepreneurship.
Dr. Sylwia Klińska works at the Department of Plant Biochemistry at the Institute of Biotechnology of the University of Gdańsk and the Medical University of Gdańsk. The team's main interests are the biochemistry and biotechnology of plant lipids. Research is focused on the search for genes associated with lipid metabolism and the determination of the biochemical and physiological functions of proteins encoded by them.
What we learned:
Dr. Klińska has experience in her research work with the Golden Gate cloning procedure, which we wanted to use in our project. She provided us with substantive support and valuable tips to help us adapt this technique to the needs of our project. She also drew our attention to aspects related to the cloning procedure - she suggested that we increase the amount of enzyme and use individual enzymes instead of a commercial mix, and extend the reaction time, which contributed to the increased efficiency of the process.
Igor Grochowina is a member of the Department of Evolutionary Biochemistry. The research conducted in the laboratory focuses on understanding the molecular mechanisms of the functioning of caregiver proteins in the most important cellular processes, as well as understanding the evolutionary mechanisms that led to the functional differentiation and specialization of caregiver proteins. Igor Grochowina has extensive experience in working with proteins, and therefore our research team collaborated with him at many stages of the project.
WHAT WE LEARNED:
Due to his experience in working with proteins, Igor Grochowina helped us to develop a plan of experiments related to testing the enzymes we selected and creating an immobilization platform - he suggested that we focus on demonstrating the activity of enzymes. Additionally, he provided us with substantive support and consultations in terms of verifying the correctness of the sequences we selected, and helped us to predict the structure of our fusion proteins using the AlphaFold2 software, providing us with their molecular models.
iGEM StartUps: Summer School was an online event that took place on 15-16.07.2023 online. The summer school was attended by a lot of students - members of other iGEM teams. We were very impressed by the sheer amount of knowledge that was shared over the two days: a lecture on IP, a discussion panel with the CEOs of their own startups, and an overview of many interesting iGEM projects demonstrating business spirit. We had the opportunity to listen to very thought-provoking presentations and participate in inspiring discussions.
WHAT WE LEARNED:
We learned what criteria the project, submitted for the Best Entrepreneurship Special Prize, should meet. An in-depth analysis of a number of exemplary igem projects showed us what steps we should still take to increase the chance of turning our idea into a future startup. We learned about the existence of many important analyses.
In addition, the lecture on IP helped us understand the relevance of this aspect, and in the future we will definitely consider submitting a provisional IP application in the initial development phase of an innovative project.
The interview with representatives of startups helped us understand the path from idea to business and gave us many valuable tips for the future.
The time to exchange thoughts with other summer school participants gave us a chance to meet Dimitrios - a member of the Thessaly 2023 iGEM team, with whom we met to talk later - you can read about our discussion below.
Spark* Gdańsk is an organization that was founded on the initiative of Spark NTNU, which operates in Norway. It offers free business support services for students and mentoring in the establishment of start-ups in a student-to-student system. The main goal of the organization is to provide substantive support in the implementation of ideas for the creation of a start-up and to provide tools and necessary training that enable young aspiring entrepreneurs to achieve their goals.
WHAT WE LEARNED:
Our collaboration with representatives of Spark* Gdańsk consisted of training under the guidance of a mentor, during which our team gained knowledge about the initial stages of creating a start-up. We learned and together with experts created basic business analysis models - SWOT, Business Model Canvas and PESTEL. Our answers were analyzed and verified by experts from Spark* Gdansk, which had a significant impact on the shaping of the Entrepreneurship strategy.
A representative of our team took part in science popularization workshop, which took place on 20/05/2023 at the Medical University of Gdansk. The workshop was led by Dariusz Aksamit, a medical physicist associated with the Department of Nuclear Physics at the Warsaw University of Technology, the "March for Science" foundation and the "Advocates for Science" association. Dariusz Aksamit has been involved in the popularization of science for years, conducting trainings, workshops and appearing in the media as an expert. During the workshop he answered an important question: how to talk about science with those who are not scientists? He shared a lot of practical advice on conducting workshops, pointed out the most common mistakes in communicating with interlocutors and hinted at how to avoid them. The presenter repeatedly stressed that a good science popularizer is characterized by openness to constructive criticism, humility and willingness to communicate. It is the communication between presenter and participants that is crucial, because workshops should be a dialogue, not a monologue. The presenter also pointed out that the most important question when working on a project should always be: "so what?". How can we make a change? And why should others care about our project? The workshop with Dariusz Aksamit was a unique opportunity to learn the secrets of science popularization. Without a doubt, participation in the workshop made us think a lot, inspired and motivated us to expand the popular science activities of our team. We are sure that thanks to this workshop we will develop our activities more consciously, **teaching, but also constantly learning from our audience.**
On the 15th of March we had a chance to talk with Madeleine and Theo - members of the former INSA Lyon iGEM 2022 team (France). Given that this is our first participation in the iGEM competition, the experience they shared was very valuable to us and greatly enlightened us on how the competition works. They were kind enough to answer many of our questions related to project development, fundraising, building the wiki and presenting our project. After the meeting we were able to look more broadly at the challenge ahead of us and sat down to plan the tasks that awaited us over the coming months. Tips from Madeleine and Theo helped us immediately during the fundraising phase.
In June, we found out that a member of the current INSA ENS Lyon1 2023 team (France), Maelys, was staying in Poland for a while. We are very happy that we were able to meet in beautiful Gdansk and talk about our teams' projects. We shared our fundraising and team formation experiences. Due to the fact that Maelys is responsible for educational activities in her team we exchanged ideas about social media activities and how to promote the project. It was wonderful to get to know your perspective Maelys!
In July, we encountered difficulties ordering DNA sequences from IDT and TWIST. Grzegorz and Anton - members of Lund Team (Sweden) came to help us. They told us how their team prepared the sequences for ordering and discussed the problems we encountered. With their help, we were able to quickly prepare the order and get to work. We also had time to talk about our projects, which both involve bioremediation. We discussed the possibility of implementation of our solutions into the Wastewater treatment plants.
Seoul-Korea team members reached out to us to talk about our projects and exchange iGEM experiences. Our projects share a common problem of water pollution, but they involve quite different techniques. Their project centers around biosensors, which was a completely new topic for us. On the other hand they were curious about our cell-free approach and the production of enzymes for water purification. The whole conversation was very intriguing and certainly brought to our attention the need for sensors for water analysis.
In August, we had the opportunity to speak with members of the Thessaly (Greece) team - Dimitrios and Argyrios, whom we met during iGEM StartUps: Summer School. Both of our projects touch on bioremediation, so it was a great opportunity to compare our approaches to the topic. Although our meeting was mainly dedicated to Entrepreneuship and Education parts of the projects. Discussing our ideas allowed us to see what analysis we can still do and what actions we can take to further inspire others with a passion for science.
Our actions taken as part of the iGEM 2023 competition focused on solving a niche problem that has an impact on the quality of life and health of the population on a global scale. During the project, we gradually raised awareness among people about the dangers of active compounds, including phthalates. We presented our ideas to potential users and asked about their expectations, responding to all suggestions by introducing appropriate changes in response to these expectations. Our actions were supported by numerous consultations and discussions with experts who provided us with factual information confirming or disproving our ideas. All of this made it possible for us to create something that would be responsible and good for the world. Initially, we planned to create a bacterial filter to remove phthalates from water. However, our first contact with the community revealed that this idea needed to be improved, due to public reluctance to drink water from a bacterial filter. This gave us a signal to modify our plan in a way that would be more appealing to the majority of the community. To this end, we consulted with Prof. Michał Obuchowski and created a plan that would allow us to create a cell-free filtration system. During the consultation, we learned more about the restrictive legal regulations governing the registration of GMO products in the EU, which became another argument for changing our plan. Consultation with BIOTON made us aware that the use of systems for cleaning overproduced proteins used in pharmaceutical companies would allow us to create a product that would be completely GMO-free, making it easier to implement in the real world. A visit to a local wastewater treatment plant made us understand that our future actions should focus on raising awareness among the public, including lawmakers. We also learned that it would be worth targeting our product at industrial units involved in the processing of plastic, which is required to monitor the concentration of phthalates released during industrial processes.
The activities we undertook - the mentioned above and, equally important, all the others were an eye-opening reflection for us, which resulted in the creation of a filter design that can contribute to taking responsible care of the World we live in.
