Human practices

Overview

Human Practices is a key component of every iGEM project, including ours, as it ensures that our scientific efforts are aligned with real-world needs and address important societal challenges. It’s about integrating broader societal, environmental, and ethical considerations into our project to create solutions that are not only innovative but also impactful for diverse communities. This means our project is not just shaped by the needs of scientific audiences but by the expectations of consumers, local communities, and global stakeholders alike. By incorporating these perspectives, we ensure that our work is meaningful, responsible, and capable of creating a lasting, positive impact. Our project KlothY is focused on reshaping the fashion industry through sustainability. We are developing an innovative textile made from bacterial cellulose, an alternative that addresses major environmental challenges such as monocultures, excessive water usage, and pollution. Our goal is to not only create a new material but to offer a demand-driven and future-oriented solution for the fashion market - one that emphasizes durability, resource efficiency, and a minimal ecological footprint.

To ensure our project aligns with real-world needs, we’ve engaged with a wide range of stakeholders, including fashion school students, textile engineers, environmentally-conscious citizens, and experts in fields such as environmental synthetic biology, microbiology, and economics. Institutions like Imperial College London, Michigan State University, Hochschule Niederrhein and Heinrich Heine University have provided valuable insights that have helped shape our work, ensuring that it is both scientifically robust and socially responsible. By embracing a broad spectrum of opinions, from local communities to global experts, we are working towards a solution that not only advances scientific understanding but also creates a meaningful, lasting impact on everyday life. Our vision is to demonstrate that synthetic biology, sustainability, and fashion can come together to address global challenges, reaching far beyond laboratories and into homes and wardrobes worldwide.

Contact	Colour
Expert	blue
Industry	pink
Consumer	yellow
NGO	violet
iGEM	green

Prof. Axmann, Prof. Matuszynska and Dr.Hensel 26.01.2024

Why did we meet?

After the initial project idea was developed, we were trying to evaluate how it could be translated into a feasible project. We met with Prof. Dr. Anna Matuszyńska and Prof. Dr. Ilka Axmann to understand what are the benefits and drawbacks of microbiological approaches and which modelling approaches would benefit the project. Directly afterwards, we met with Dr. Götz Hensel, to understand how working in plant systems compares to a microbiological approach.

What did we learn?

• To produce textiles with microorganisms, our first approach was to consider either introducing elongation in cells which have physical properties comparable to cotton, for instance through high cellulose content in the cell wall, or cellulose production in already elongating cells. During the first meeting, the benefits of faster growing organisms were highlighted. The focus was also in discussing the benefits of an inducible system, for biosafety and feasibility reasons, as a constitutive expression of any genes, will pose a metabolic burden to our organisms. It was also discussed how continuous cultivation will become very expensive and complicated, and how an inducible system would allow reproduction and harvesting in separate batches.
• In the following meeting with Dr. Götz Hensel, we discussed the viability of a cotton tissue culture with him. It was considered how we could create an inducible system in which we can modify the cells, so they differentiate into a fibre, and how transformations in plants are different from microbiological methods. He recommended starting in epidermal cell cultures and confirmed biolistics would be a viable option for transformation.
• We considered how we could knock out the necessary genes and what is generally possible in plant systems.

Reflection

An important role played the limitations of the cotton system, especially regarding output and efficiency. One important point was also the time a cotton boll needs to grow, which was found to be 45 - 60 days (Bo XU et. al. 2017) When considering the desired modularity of our project, it became apparent that the flexibility we needed would best be met via the microbiological path. But we were still facing the problem of finding appropriate organisms, as both initial ideas, which were to induce either cell wall reinforcements or cell elongation to the cotton fibre level, involved extensive pathways that are not tackable within the scope of our project. This was when we got the hint to meet with Prof. Dr. Markus Pauly, who is an expert for cell wall polysaccharides.

(Bo XU et. al. 2017) doi.org/10.1016/S2095-3119(16)61566-6

Prof. Dr. Markus Pauly 12.03.2024

Why did we meet?

We were facing the issue, how we could create biodegradable textiles with microorganisms, by introducing pathways of feasible complexity. As Markus Pauly focus is polysaccharide production, he was the perfect partner to explain our approach to, and discuss alternative and complementary methods with.

What did we learn?

• It was explained that through selecting organisms, which naturally produce cellulose, the genetic modifications could be limited to those necessary, to alter the properties of the final product. As Prof. Pauly worked with a group in Canada, which used bacterial cellulose as plant cell wall substitutes, he recommended looking into organisms, which produce such. When considering the progress of previous iGEM teams, as well as the stiffness of regular mats consisting of bacterial cellulose, one key point was found to be the flexibility. We discussed how hemicelluloses intercalate with cellulose, and could therefore be an interesting molecule to combine with cellulose mats, to achieve mechanical properties, which resemble regular clothing.

Reflection

When looking for bacterial cellulose producing organisms we quickly found the genus Komagataeibacter, of which we considered Komagataeibacter xylinus. As we would therefore have a fast growing, modifiable organism that directly produces extracellular cellulose, our main problem of having to introduce elongation was fixed, as we would be able to harvest a mat directly. That cuts the necessary processing steps and the connected steps of the supply chain, while still resulting in a physically resilient fabric. Our next focus was now how to modify those fabrics so they would become suitable textiles.

PhD Ronja Catharina Immelmann 18.03.2024

Why did we meet?

Ronja is a PHD student at Markus Paulys lab. Her research revolves around the synthesis of xyloglucan, a hemicellulose in the yeast Pichia pastoris. The aim of our meeting was to understand the current state of xyloglucan synthesis research in yeasts and which undertakings were the most promising.

What did we learn?

• Ronja gave us a general introduction to xyloglucan and how it is synthesized. She outlined which proposed enzymes showed results and on which parts there is still work to be done and how she will try to solve those problems. We discussed which approaches we would use and why we saw Saccharomyces cerevisiae as the fitting choice for our project. We discussed which alternative approaches could also be established to produce xyloglucan in yeasts and which specific factors will be similar and different in a yeast like S. cerevisiae. We also explained and discussed the concept of human practices and the impact is has on the quality and applicability of research in general.
• S. cerevisiae is a promising alternative organism for expression of plant cell wall polysaccharides as xyloglucan.
• For our goals a challenge might be the final localisation of the produced xyloglucan.
• Specific xylose transporters into the Golgi apparatus are probably important for the production of a more xylosylated glucane backbone.

Reflection

As it was proposed the xyloglucan might interact with the natural cell wall of the yeast. We first thought we needed to prevent the interaction with other polysaccharides in the cell wall. After evaluating her and Markus Paulys feedback we planned on evaluating the final locations in the end, but also realized that the interaction with the cell wall might bind the yeast tight to the cellulose mat. That might result in closer interaction between the yeast and our cellulose producing bacterium Komagataeibacter xylinus. Through this closer interaction a more evenly distributed co culture might result in fabrics with a homogenous texture and color pattern. A specific UDP-xylose transporter was chosen instead of a less specific UDP-glucuronic acid transporter.

Prof. Dr. Johannes Hegemann 19.03.2024

Why did we meet?

Professor Dr. Hegemann is an expert for working with yeast, namely Saccharomyces cerevisiae. He was therefore the next point of contact, to evaluate, if our approach was feasible, and which yeast would be most fitting for our purpose.

What did we learn?

• In the meeting we discussed that S. cerevisiae is so well characterized, that for bringing a new pathway into the organism, it would be most fitting. Yarrowia lipolitica was also shortly considered for applications such as waterproof textiles, as it has a diverse lipid metabolism, but it was decided to focus the project on fewer aspects.
• We also discussed if the Golgi-transport of our hemicelluloses will work, and how they might interact with the fungal cell wall. Finally he explained different surface display systems.

Reflection

In conclusion of the meeting we settled for S. cerevisiae as our property modifying organism, next to K. xylinus as our extracellular polysaccharide producer. We also incorporated the advice regarding surface displays, for planning a hemicellulose binding domain.

Prof. Dr. Oliver Ebenhöh and Dr. St. Elmo Wilken 27.03.2024

Why did we meet?

We reached out to Professor Oliver Ebenhöh, a leading expert in mathematical modelling and the head of Institute of Quantitative and Theoretical Biology at Heinrich Heine University Duesseldorf, to gain insights and feedback on our project. Professor Ebenhöh is renowned for his innovative research in simulating and describing the production and degradation of biopolymers, including starch and complex secondary metabolites, as well as the timing of metabolic processes. Given his expertise, we sought his opinion on our initial ideas for developing a mathematical model aimed at optimising production efficiency, enhancing the properties of our biopolymer textile, and predicting the behaviour of our dye system.

What did we learn?

• Limitations of Metabolic Modelling:

  We discovered that traditional metabolic modelling techniques, such as flux balance analysis, have limited accuracy in predicting biopolymer production. This necessitates exploring alternative approaches for more precise predictions.

• Differential growth conditions with K. xylinus:

  It is a viable strategy to grow Komagataeibacter xylinus with varying conditions and employ linear regression to model changes in the properties of the biopolymer. This black-box machine learning approach can effectively predict outcomes based on observed data.

• Chromoprotein Expression:

  Modulating chromoprotein expression was highlighted as a promising area. We should investigate titratable inducers that offer a correlation between inducer concentration and protein production to fine-tune expression levels.

• CMYK Dye System:

  The simplified CMYK system for dye application was found to be an intriguing concept. Techniques such as maximum likelihood estimation can be utilised to construct a robust mathematical model for this system.

Reflection

After our conversation with Professor Oliver Ebenhöh, we refined our mathematical modelling approach to focus on the most promising models. We also consulted with our secondary PI, Dr. St. Elmo Wilken, regarding titratable inducers and discovered a promising paper on a four-molecule inducible system in Saccharomyces cerevisiae, utilising vanillic acid, xylose, aTc, and IPTG (Park et al., 2023). This system is ideal for our purposes.

Additionally, we brainstormed various growth conditions to modulate. We decided to experiment with altering the concentration of xyloglucan. We hypothesised it will intercalate with the bacterial cellulose matrix and modify properties of the mat. (See Property Test) We also considered adjusting the concentration of K. xylinus' carbon source, glycerol, to optimise production conditions.

Prof. Dr. Daniel C. Ducat 03.04.2024

Why did we meet?

The central part of our project is the co-culture between Saccharomyces cerevisiae and Komagataeibacter xylinus. Because stable co-cultures are quite difficult to establish, we wanted to pinpoint some variables we can focus on to optimise our approach. Furthermore another idea of ours was to introduce a third party into the co-culture, namely cyanobacteria, which would produce a carbon source to feed the other microbes in the coculture and therefore make it more sustainable. For this we contacted professor Daniel C. Ducat who is a biochemistry and biotechnology professor at Michigan State University that specialises in co-cultures especially those containing cyanobacteria.

What did we learn?

• Daniel Ducat advised us to perform a metabolic modelling of both organisms before establishing a co-culture to have an estimation of the behaviour. Furthermore, he suggested we work on two variables in the cultivation process to get an understanding of the influence they have.
• He also suggested physical as well as chemical property testing to us for example tensile strength tests of our produced cloth.
• Further, Daniel Ducat underlined the importance of a Life cycle analysis to get an understanding of the viability of our product, for which he recommended professor Oliver Ebenhoeh from the Heinrich Heine University.
• In the long run, we also discussed the possibility of introducing cyanobacteria into our co-culture to have an alternative carbon source to make the whole production process more sustainable.

Reflection

Because of this meeting we started working on metabolic modelling especially for the knockouts and knockins we planned for the bacterial cellulose synthesis complex. Furthermore, it validated our plan to do mechanical property testing, which we later did at the Niederrhein University of Applied Sciences and compared the results of wildtype produced mats to mats where xyloglucan was added beforehand in the media. Daniel Ducat also emphasised the importance of a Life Cycle analysis (LCA) which proved to be one of the central points that occurred in meetings with industry and stakeholders. To assess this LCA we also planned to reduce the costs for cultivation by utilising the photosynthetic properties of cyanobacteria, which would save resources and make the resulting product cheaper. Furthermore the two variables we chose were sugar and temperature, we conducted cultivation experiments where we varied those two factors to see how the behaviour of the microbes change. For example it appears K.xylinus forms a mat way easier in standing cultivation on RT then on 30 degrees in an incubator.

Independent Researcher Sebastian S. Cocioba 08.04.2024

Why did we meet?

Sebastian Cocioba is an independent researcher who specialises in flower design and making biology accessible. He embodies the iGEM spirit, researching independently, struggling and learning from mistakes. Since he is very knowledgeable across a wide range of topics as well as chromoproteins which play a vital role in our project, we wanted to talk to him to gain insights on how to approach our project, how to deal with failure and how to approach our CMY chromoprotein system.

What did we learn?

• We developed the idea to conduct “colormixing” experiments with bare chromoproteins to see how the colours interact and base a software on it.
• Glycerol can act as a moisturiser to keep the proteins stable for longer or alter the properties of the bacterial cellulose mats.
• Future work on a melanin binding domain to attach pigments (for example melanin) directly to the cellulose fibres could be viable.
• For a true CMYK system we should look at Tyrosine Melanin expression as black colour.

Reflection

The meeting was invaluable to kick off our project and to learn a lot about various topics. While the main focus was on the dyeing of our bacterial cellulose mats through chromoproteins, including on how we choose the ideal chromoproteins to form our colour palette through fPBase.org and how we should look into a separate implementation of the colour black as equal saturation of cyan-, magenta and yellow chromoproteins would not be sufficient. Sebastian went even further and actually gave us valuable insight which not only helped us shape our cloning strategy for the dye part later on but the project in general. For instance, he provided valuable feedback to our initial idea to use hemicellulose to increase flexibility for and gave us alternative options to try should certain parts of our project not work. One example being, to use glycerol post treatment as an alternative to hemicellulose to increase flexibility if our goal should fail. Furthermore, he was also advocating to look into creating a hemicellulose binding domain (HBD) to connect the chromoproteins, instead of the cellulose binding domain (CBD) and do comparative experiments for our dyeing part. Sadly, we could not implement all of Sebastian's feedback into our project due to some arising challenges in our project. However, he really inspired us to really look into our project and the challenges it entails and we are thankful for his support.

Prof. Dr.Piergiuseppe Morone 09.04.2024

Why did we meet?

For better understanding of the current trends in the textile supply chains and their impact on the current textile market, we were talking to economic experts, who can give us better insights. Fortunately for us, one of our advisors had an ideal contact ready: Prof. Dr. Piergiuseppe Morone, A professor of Economic Policy at Unitelma Sapienza in Rome, Italy. He is an expert in researching economic trends in the textile industry, with a strong interest in green innovation and sustainable circular bioeconomy. Furthermore, he is advocating for the transition from current fast fashion trends to a more sustainable, circular, economic system. Because of this, we contacted him to discuss the current failings of the textile industry and market and why he thinks that a change in consumer behaviour is necessary. Additionally, we were also interested in his opinion on our project idea and which physical properties our product may need to fulfil to become more sustainable.

What did we learn?

• Prof. Morone identified the fabrication stage as having the greatest potential for reducing environmental impact. Sustainable practices here, such as using organic cotton and innovative synthetic fibre technologies, can significantly lower resource use and emissions currently produced by industry
• Prof. Morone believes our product can make a difference, emphasising its biodegradability. However, he also mentioned that fabrics need to be durable for a long duration, reducing the need for frequent replacements due to weak material integrity common in fast fashion products, as we would otherwise fall into the danger of becoming another option to produce fast fashion, destroying some of our efforts to achieve a sustainable product.
• In terms of how to convince potential customers of the viability of our product. He suggested that educating consumers through tangible evidence of our product's benefits is crucial.
• Many consumers are unaware of their environmental impact, and demonstrating the advantages of sustainable practices over traditional methods can drive behaviour change in his opinion.
• Specifically conscious consumers are more likely to choose our product if we transparently showcase its safety and environmental benefits.
• Key sustainability assessment parameters include fabric durability, biodegradability, reducing the need for mixed fibre fabrics, and addressing the general issues of synthetic fibre.
• Morone finally mentioned that our next steps should involve showcasing our product's sustainability, developing educational materials, and gathering and generating comparative data. (See LCA)

Reflection

During our discussion with Prof. Morone, we gained new insight on how the fashion market behaves and actually left the meeting with a more grounded way on how we should go about the product. While we were glad that Prof. Morone thinks that our product could actually make a difference, especially after explaining how biodegradable our material is compared to blended materials. We became more sensitive to one of our perceived strengths. One of them being that compared to cotton to make our product more sustainable.which needs to grow for 160 days before it can even be harvested and refined, our bacterial cellulose mats can be extracted after just 8-14 days. We were beginning to understand that being a faster method or using a sustainable carbon source aren’t the only determining factor for sustainability and may even be a detriment in achieving a true sustainable product. Since we would just by further fueling fast fashion text. We were therefore committed to better understand how durable our material really is and how we could potentially improve it. Furthermore, while we were also then further committed to achieve an initial LCA model to be able to roughly compare our product more with current textile options, Prof. Morone really emphasised that in fashion, buying second use is always more sustainable than buying a new product. Therefore the best thing we could really achieve is creating a product that is firstly, using sustainable carbon sources and secondly, durable enough to be used for multiple generations and thirdly can potentially be broken down further to create new textiles. Achieving a true circular economy in every sense.(See iGEM Dresden)

Dr. Amanda Staudt 22.04.2024

Why did we meet?

We reached out to the Leibniz Institute for Interactive Materials, a prestigious institution dedicated to advancing materials that integrate technology and nature - a vision that very much resonates with us and our project. The institute aims to enhance materials' ability to adapt their properties to changing external conditions. We had the opportunity to speak with Dr. Amanda Staudt, a project manager in biotechnology at the institute. Our discussion focused on various property testing methods that could be applied to our bacterial cellulose (BC) pellicles.

What did we learn?

• Mechanical Testing:

  Conducting mechanical tests, such as tensile strength and Martindale abrasion tests, will be crucial to evaluate the robustness of our material.

• Chemical Testing:

  Performing chemical tests, including flammability tests, is essential to ensure that our textile is safe for use as clothing.

• Cultivation of K. xylinus:

  It's important to set the dimensions of the BC pellicle appropriately during cultivation to obtain meaningful and consistent results.

• Predictive Mathematical Modeling:

  When building a predictive mathematical model based on property tests, it's vital to ensure that tests are replicated to achieve reliable and accurate predictions.

Reflection

After our conversation with Dr. Amanda Staudt, our experimental design for both cultivation and the property testing became much clearer, including the specific container dimensions needed. We also gained a better understanding of defining cultivation parameters, such as different hemicellulose concentrations, and assess the properties with linear regression.

Dr. Staudt recommended six replicates for the tensile strength tests and three for the Martindale abrasion tests. She specified that the dimensions for the tensile strength samples should be 6 x 2 cm, and the diameter for the Martindale abrasion samples should be larger than 50 mm.

Based on these recommendations, we calculated the necessary volumes of chemicals required to produce the BC pellicles. These pellicles were then tested on site at the Niederrhein University of Applied Sciences. (See Property Testing)

iGEM Team Dresden 26.04.2024

Why did we meet?

One of our constant concerns especially after the meeting with Professor Morone is sustainability and how to make sure that the textile we produce does not end up in landfills just like all the blended materials that are currently used. Because of this we looked at other iGEM teams projects, because we wanted to engage with young ambitious scientists that want to tackle the current crises and may even be the missing puzzle piece to bring our idea to a circular economy. Not long after initiating the search we found the iGEM Team Dresden “ReFiBa” who focus all around genetically engineering Bacillus subtilis to produce cellulase and PETase and immobilise these enzymes on the surface of the bacillus endospore. The enzymes then break down blended materials as well as pure cotton. The cotton part will get broken down into sugars. Our idea was to start a collaboration to see if iGEM Dresden hydrolyses our bacterial cellulose mats and if successful we can use the resulting product as a carbon source to grow new bacterial cellulose mats. Achieving a transformation from difficult to recycle material to an easier recyclable material.

Reflection

The collaboration with iGEM Dresden is a really interesting opportunity for us and our project. With this we are able to come closer to a circular economy like Professor Morone also later suggested. Furthermore this would also decrease the price of our textile which would open up new potential customers as Roland Stelzer suggested. Also another upside would be to tackle the massive landfills and introduce textiles that are easier to recycle. In the beginning of September we made the collaboration real but due to time constraints the data will be collected after the wiki freeze.

Dr. Dipl.-Ing. Sascha Schriever 29.04.2024

Why did we meet?

We reached out to the Institute for Textile Technology of RWTH Aachen (ITA), a prestigious institution dedicated to the holistic biotransformation of textile technology through biological principles for small-scale value-added processes. One of their notable projects is the BioSC's LignoTex project, which aims to enhance the economic viability and reduce the CO2 footprint of biorefineries by developing a comprehensive process for extracting, modifying, and spinning lignin from various plant sources into fibers for textile applications. We had the opportunity to speak with Dr. Sascha Schriever, head of Chemical Technologies for Fibre and Textile Innovations, and Dr. Roshan Paul, Senior Program Manager. Our discussion focused on improving the property performance of our bacterial cellulose (BC).

What did we learn?

• Chemical Treatment:

  post-processing of BC with chemicals such as Perfluoroalkyl and Polyfluoroalkyl Substances (PFAS) may enhance its hydrophobicity, however due to its persistence in the environment, ITA is developing bio-based alternatives.

• Purity Analysis:

  Analysing the cellulose purity in BC pellicles could be beneficial, as their research indicates that removing impurities like lignin and hemicellulose results in stronger fibers.

• Solubilising and Spinning:

  Investigating the process of solubilising BC and converting it into fibers and yarns might be worthwhile if optimising tensile strength is the primary goal

Reflection

Following our discussion with Dr. Sascha Schriever and Roshan Paul, we expanded the scope of properties to be tested, considering hydrophobicity as an additional dimension. Enhancing hydrophobicity could be valuable, particularly for producing textiles tailored for water-related activities where waterproof properties are essential.

Although our approach aims to create a ready-made textile "mat" to circumvent downstream processes like solubilisation and spinning, we found their work and approach originating from a plant-based system fascinating and insightful.

Prof. Dr. Tom Ellis 03.05.2024

Why did we meet?

Through the early researching stages of our project we discovered a name often named in papers connected to bacterial cellulose and Komagataeibacter and two paper specifically caught our eyes as one was discussing a co-culture between Komagataeibacter rhaeticus and Saccharomyces cerevisiae and the other was discussing the viability

(“Self-pigmenting textiles grown from cellulose-producing bacteria with engineered tyrosinase expression”).

(“Living materials with programmable functionalities grown from engineered microbial co-cultures”).

Both papers are co-authored by Tom Ellis, an esteemed professor for synthetic genome engineering from the Imperial college of London. We established contact with him to learn more on what different factors may affect our co-culture, for example which carbon source we should use and why, how we can make sure that one organism does not outgrow the other. Also since the Komagataeibacter strains are not model organisms, transformations can pose quite a challenge and because our goal is to make the bacterial cellulose synthesis inducible we wanted to get his opinion on our approach and the viability because he was successful with transformation.

What did we learn?

• Tom Ellis recommended the use of YPS Media in our co-culture since K. rhaeticus (the strain he uses) can’t metabolise the sucrose easily but S.cerevisiae can so a slight codependency is established.
• He reaffirmed that chromoproteins are only viable as a proof of concept with no real life application. Since they get damaged or destroyed easily. Heat, drying and proteases (found in laundry detergent) destroys them and makes them lose their colour.
• Pigments would in his opinion allow a real CMYK system since there are black pigments like eu-melanin and are more tolerant to heat and dry conditions. Komagataeibacter strains cannot be incubated in a shaking incubation unless cellulase is added. Because the shaking results in cellulose strings through the whole tube instead of pellicle formation. Furthermore, it also leads to the bacteria mutating frequently.
• A knockout or inserts of parts of the bacterial synthesis operon is extremely hard and has not been accomplished yet.

Reflection

Tom Ellis validated that chromoproteins are only a proof of concept for our project and acts as model based on it due to their inherent instability but they still can lay the foundation for possible future application in the textile industry, where the chromoproteins are switched with heat stable bacterial pigments, which would not have the same longevity problems as chromoproteins. Furthermore this modular approach could have a vast impact on the textile industry since the dying process would occur while the mat grows and because of this and the natural produced pigments reduce harmful chemical use. To test if bacterial pigments or natural pigments in general would be viable we decided to integrate experiments with pigments in our lab work using curcumin, bacterial produced violacein and beta carotene. Furthermore, due to the suggestion of a different carbon source in our cultivation media (at this point we only used dextrose) we decided to test carbon sources like glycerol, sucrose. Not only did we discover that K.xylinus can metabolise sucrose really well, it also changes the properties of the forming bacterial cellulose-Mat. For example YPS mats are almost transparent while mats grown on YPD look like parchment paper. Tom Ellis critique and insight in the function and behaviour of Komagataeibacter was vital for the project, furthermore his critique motivated us to further broaden our scope and experiment more. Although Tom Ellis warned us that a knockout strain is hard to achieve and the transformation of K.xylinus in general is quite difficult we still decided to pursue this experiment because it would greatly impact our project and the scientific world as a whole.

Prof. Dr. Daniel C. Ducat 10.05.2024

Why did we meet?

After further researching co-cultures as well as talking to Tom Ellis we were looking for some practical advice on how to investigate our co-culture, as one of the biggest challenges for us was how to properly quantify how to stable a K. xylinus and Saccharomyces cerevisiae co-culture really is. For this we contacted Prof. Dr. Daniel C. Ducat once more to learn more of all the potential methods of assessing the quality of a co-culture and how we could solve some of the challenges we have found in during, including that K. xylinus prefers to stay at the liquid air barrier for maximum oxygen uptake during standing cultivation meanwhile Saccharomyces cerevisiae tends to collect at the bottom, which would result in an uneven distribution and incorporation later in an inducible system.

What did we learn?

• Measuring growth in liquid cultures is easier than on plates, since the cultures are more homogenous. However due to the cellulose formation OD-measurement of viable cells would be insufficient. And transforming one of the partners with a fluorescent marker and measuring relative fluorescent units may be a decent alternative.
• Some of the yeast cells may get trapped between the forming cellulose fibres which would be beneficial because the metabolites we want the yeast to produce would also be between the cellulose fibres.
• He advised us to find a way to increase the yeast cell concentration by either engineering the yeast to actually be able to increase concentration or somehow trap it reliably between the cellulose fibres.
• He advised we should focus on the bacterial cellulose mat to quantify the yeast in it, since this is the place where it matters most. For this, he suggested using fluorescent, GFP-expressing yeast to locate it on/in the cellulose.
• We talked about using sucrose as the carbon source for the co culture because of the reasons Tom ellis mentioned (K. rhaeticus can not metabolise sucrose easy but yeast can)
• He suggested performing a qPCR for quantifying the DNA quantities to get a rough estimate of the co-culture ratios.

Reflection

Due to talking with Daniel Ducat about making the yeast “sticky” we further researched this topic and ended up designing a part where a cellulose binding domain is attached to an anchor protein which is displayed on the surface of the yeast. This also led to us using another Saccharomyces cerevisiae strain namely EBY100 due to it being engineered to be able to display heterologous proteins the beforehand mentioned surface display protein. Furthermore we succeeded in cloning and transforming this part. As already mentioned in the meeting with Tom Ellis we ended up testing different carbon sources especially sucrose to see if and how well K.xylinus can metabolise it, we discovered contrary to K.rhaeticus, K.xylinus does grow really well on sucrose media and the forming mat is way more transparent which also makes it preferable for dyeing.

Fashion Student: Ann-Kathrin Wasow 31.05.2024

Why did we meet?

We met with Ann-Kathrin Wasow because of her deep understanding of textiles and her focus on material innovation within fashion. As a fashion student, she has dedicated her work to exploring both the aesthetic and functional aspects of fabrics. Her insights were particularly valuable for our project, as she brings a fresh perspective on how sustainable materials can not only be eco-friendly but also align with the needs of modern consumers. Her experience with different types of fabrics and her expertise in sustainable design made her a perfect collaborator to help refine our approach.

What did we learn?

• Ann-Kathrin provided us with valuable insights that challenged our thinking about fabric choice. She explained that natural fabrics, like cotton or linen, are often preferred over synthetic ones because they feel much more comfortable and breathable on the skin.
• She also emphasized that clothing isn’t just about functionality - it’s a reflection of personal style and character, which means that the materials we use need to create a positive sensory and emotional experience for the wearer.
• Additionally, she highlighted a crucial point: while consumers are increasingly drawn to sustainable fashion, the high price of these garments often drives them away. Ann-Kathrin told us that if we want to make a real difference with our eco-friendly materials, we must ensure that our production process is cost-effective to make the final product affordable.

Reflection

Our conversation with Ann-Kathrin made it clear that we need to think beyond just the durability and sustainability of our material. She made us realize that in order to succeed in the competitive fashion market, our material must also deliver on comfort and wearability. It should be light, breathable, and soft, without any roughness or stiffness that might turn people away. Moreover, versatility is key - our material must be adaptable enough for various clothing designs, and resilient enough to withstand daily wear. This feedback pushes us to carefully consider not only the ecological footprint of our material but also how it feels and functions in everyday use, so that it can truly stand out as an appealing option in the world of sustainable fashion.

Non-Governmental Organization: Global Organic Textile Standard (GOTS) 06.06.2024

Why did we meet?

As our project focuses on developing a product produced by GMOs, it is important for us to learn about all perspectives, including more critical ones. For that, we contacted Global Organic Textile Standard (GOTS). Global Standard NGO (the organisation responsible for setting and establishing the GOTS standard) is a non-governmental organization that focuses on the development, promotion, protection and implementation of the Global Organic Textile Standard, an international standard for the processing of organic textiles along the entire supply chain. and certifying companies that comply with those standards already. Companies involved in the first processing step of organic fibres, over production, processing and manufacturing until the final step can be certified to GOTS, given they meet the stringent criteria. They are advocating for the use of sustainable and environmentally friendly harvesting methods (organic) of natural fibers, worker's safety, fair compensation, environmentally-friendly production and sustainable business practices. As many of their goals align with ours, we felt they were the perfect candidate to have a fair discussion about the issues of the current textile market, what in their view are the best possible steps to alleviate those issues could be, and finally their opinion on GMOs in general and products produced by GMOs are. This led us to talk to Juliane Ziegler who is the Representative in Germany, Austria and Switzerland of the GOTS.

What did we learn?

• Juliane Ziegler described our project as: "it's an interesting pilot" , "sounds like an idea definitely worth exploring"
• We need to conduct a life cycle analysis (LCA) to ensure that the product we produce is economically viable
• Currently we would not be able to receive the GOTS sigil since the usage of GMOs is not allowed in their requirements as their focus is on organic textile production.
• NGOs are important to keep companies accountable for their actions, enforcing global standards and communicating their implementation to the public.

Reflection

We were happy to hear that even though the NGOs and our view on the usage of GMOs for textile production differ, our goals to pursue an ethic and sustainable production align and we were able to convince them of our idea in a sense that we aim for a more sustainable, regionally independent approach from the normal textile production. Furthermore, despite not complying with the current set standards of GOTS, we still believe we could one day still produce ethically and locally as well despite our material not classified as "organic textile". So after the meeting we followed their advice and did experiments to be able to achieve a rudimentary version of an LCA. Unsurprisingly, we discovered that the price of our product would currently be about 30 times as expensive as regular cotton. But as we were expecting this to be the case, we were already putting effort to bring the economic scale more in our favour by researching and working on solutions to make our textile more cost efficient (see iGEM Dresden and iBC). This also shows that if we want our solution to be an actual method of textile production, we would need to cooperate with a company who is actively looking for alternatives, new experimental textiles. This led to us to later establish contact with Roland Stelzer (meeting link) to learn more about a company's perspective on our project.

Sustainability Day 05.06.2024

Why did we meet?

We had the opportunity to showcase our project KlothY at a booth during the Sustainability Day, an event organized by the Heine Center for Sustainable Development – Diversity, Environment, and Health (HCSD). The reason we wanted to present our project at this event was clear: the entire day revolved around the theme of sustainability. With a diverse array of projects and booths all dedicated to sustainable initiatives, we knew that our project aligned perfectly with the event’s goals. Our aim was to leverage this setting not only to raise awareness about KlothY, but also to gather valuable feedback from the audience and fellow exhibitors. This time, however, we focused on gauging reactions from a purely sustainability-driven perspective rather than from a scientific or technical standpoint. We wanted to understand how people perceive a synthetic biology project like ours within the context of sustainability.

What did we learn?

• Although a major part of our involvement at Sustainability Day was presenting KlothY to visitors, the event offered us several important lessons. The most striking takeaway was related to the public image of our synthetic biology project. For many people, sustainability is more of a "green" image, often associated with nature and minimal technology, rather than a concept that can include synthetic biology or genetically modified organisms (GMOs). While the audience at Sustainability Day was already inclined towards sustainable initiatives, many were surprised and intrigued by the idea that a project they perceived as purely “GMO-based” could also be sustainable. This led us to a critical realization: synthetic biology and other STEM fields, particularly those involving genetic engineering, have a significant image problem when it comes to public perception.
• We learned that projects in synthetic biology, including ours, need to work on how they are portrayed to reach a broader audience. The disconnect between the idea of “synthetic” and “sustainable” was clear, and overcoming this perception barrier is essential for greater acceptance and understanding of bioengineered solutions in sustainable development.

Reflection

The feedback we received for KlothY reaffirmed the growing interest and demand for sustainable alternatives to conventional fashion production. However, it also highlighted a considerable gap in how we communicate the sustainability of synthetic biology projects to the public. Many people, it seems, ar e lost to the field of synthetic biology because of the negative connotations tied to its image. While this revelation didn’t necessitate changes in the technical execution of our project, it had a profound impact on how we plan to present ourselves in the future.

One notable draw to our booth was the prototype of our property testing machine, designed and built by our team member Noah Ben Bulawa. The novelty and uniqueness of the device were key factors that captured visitors' attention and sparked curiosity. This fascination underscored the importance of continuing to engage audiences with innovative and visually striking elements. Moving forward, we realized that tapping into this sense of curiosity and wonder is crucial for generating interest in synthetic biology and for shifting public perception. We plan to build on this fascination, using it as a gateway to introduce people to the sustainable potential of our work .

Dr. Ir. Pieter Nachtergaele 19.06.2024

Why did we meet?

As part of our mission to enhance the sustainability of textile production via bacterial cellulose (BC), we sought guidance on Life Cycle Assessment (LCA) from an expert with a deep understanding of renewable resources in the chemical and packaging industries. We met with Dr. Pieter Nachtergaele, a postdoctoral researcher at the Department Green Chemistry & Technology at Ghent University. He is also Bioeconomy Youth Ambassador at the European Commission with a passion for using process systems engineering tools to advance the transition to a bio-based and circular economy. His expertise in assessing and improving the sustainability of technological operations, food systems, and packaging recycling made him an invaluable contact for our project.

What did we learn?

• Carbon Source:

  Dr. Nachtergaele emphasised that the choice of carbon source is the most crucial factor in determining the economic viability of our process. This insight will guide our approach as we refine our methodologies and aim for a more sustainable and efficient production process.

• Circular economy:

  He was supportive of our collaboration with iGEM Dresden in reusing depolymerised fabrics as the substrate for K. xylinus to produce textiles.

• LCA principles:

  He introduced us to paid and open-source LCA software and databases that we can use to get predictive numbers in formulating an LCA model.

Reflection

Following our discussion with Dr. Pieter Nachtergaele, we recognised the need to decide on the carbon source we will use to produce our BC mats. To this end, we will experiment with media with different carbon to grow BC mats (See Niederrhein Tests under Property Test)< and communicate with the iGEM Dresden team about the different types of substrates that can be extracted from their process and explore the potential integration with our BC mat production.

After surveying all institutes at our university for licensing and guidance on LCA, we found that conducting a full LCA is currently beyond our scope. Instead, we have decided to focus on determining key performance indicators of our process, specifically yield and rate of production. These metrics will guide our efforts in refining our production approach and ensuring economic viability. You can read our experiments page for more. (See LCA)

Modedesign-College Duesseldorf 10.07.2024

Why did we meet?

The Mode Design College (MDC) is a prestigious institution known for shaping the future of international fashion design, communication, and graphic design. Our iGEM team saw MDC as the perfect partner to present our project, KlothY, which is part of the iGEM Village focused on Fashion & Cosmetics. We wanted to bridge the gap between synthetic biology and fashion design, and showcase how biotechnology can revolutionize the textile industry, offering new opportunities for sustainable innovation in fashion.

Our contact with MDC was established because we recognized the importance of integrating scientific advancements with the fashion industry. By introducing KlothY, a textile made from bacterial cellulose and hemicellulose, we aimed to demonstrate the potential of biofabrication in creating eco-friendly materials. Our goal was to show that biotechnology can contribute to reducing the environmental footprint of fashion by developing more sustainable production methods. This visit was an opportunity to engage students and faculty in a discussion about the impact of biotechnology on fashion and how it could address some of the industry's pressing sustainability challenges.

What did we learn?

• During our visit, we introduced MDC students and faculty to the basic concepts of biotechnology and synthetic biology within the iGEM framework. Our focus was on the unique characteristics of bacterial cellulose and hemicellulose, particularly how these materials could be used to produce eco-friendly textiles. This introductory session was key in helping the students understand the potential applications of synthetic biology in fashion and how it could play a role in transforming the industry toward more sustainable practices. By highlighting the environmental benefits of developing biodegradable and renewable materials, we set the stage for a productive exchange of ideas. After our presentation, we encouraged the students to brainstorm their own synthetic biology-based fashion ideas. In this open-ended, creative session, students were free to explore and push the boundaries of how biological principles could be applied to fashion design. This exercise led to a variety of innovative and imaginative concepts, showcasing the students' ability to think creatively and consider new possibilities for the future of fashion. Their ideas helped broaden our perspective on how KlothY and similar biotechnological innovations could be applied in ways we had not previously considered.

• We also provided the students with samples of KlothY, asking them to evaluate and critique it from a design perspective. This hands-on experience was incredibly valuable for us, as it allowed the students to assess the material's potential in terms of aesthetics, practicality, and how it could be integrated into existing fashion processes. The feedback we received was highly constructive. Some students suggested using KlothY in high fashion (haute couture) designs, while others offered practical advice on how to improve the material's texture, flexibility, and durability. The diversity of feedback highlighted the versatility of KlothY and offered us new insights into how to further refine the material for broader applications in the fashion industry.

Reflection

The feedback and ideas we received during our workshop with MDC were instrumental in refining our KlothY project. The suggestions from the students and faculty helped us shift our focus to key aspects such as improving the texture and durability of the material. These practical insights pushed us to consider how KlothY could be adapted to meet the real-world needs of fashion designers and consumers alike. This collaboration also opened up new possibilities for integrating biotechnology into fashion, making us rethink how our material could fit within both niche and mainstream fashion markets.

The collaboration with MDC clearly demonstrated the value of interdisciplinary cooperation. By merging scientific innovation with the creativity of fashion design, we were able to explore new directions for the development and application of KlothY. The students' fresh perspectives and the faculty's expertise guided us to not only enhance the quality and usability of our material but also explore innovative ways to make it more appealing for a wide range of fashion applications. This experience reinforced the importance of considering both scientific and creative viewpoints to ensure that biotechnological advancements can truly make an impact in industries like fashion.

Ultimately, this workshop highlighted how scientific innovation can be enriched through collaboration with creative fields. It provided us with a clearer vision of how biotechnology can shape the future of sustainable fashion. We now feel more confident that KlothY can be adapted to meet the demands of the fashion industry, opening the door to new possibilities for environmentally friendly and innovative textiles.

„If you stick to organic dyeing materials, the seamless growth process and missing processing steps will certainly be a huge improvement in the production of eco-friendly textiles. I mean, I’ll certainly wear it.“ - Verena Schamp, textile engineer and lecturer at MDC.

Hochschule Niederrhein: M. Sc. Mark Neumann 10.09.2024

Why did we meet?

We wanted to learn more about the possible applications and the mechanical properties of our material to really understand what specific properties we need to work on. Furthermore we wanted to use professional equipment that tests by standardised protocols to ensure accurate data and comparability with current textiles. For this we contacted Marc Neumann of the University of Applied Sciences Niederrhein from the department Faculty of Textile and Clothing Technology. We wanted to learn about possible tests we could perform as well as the requirements and preparations needed to perform those.

What did we learn?

• Our material has high tensile strength (relative comparing top standard clothing articles (DTB))
• Due to bacterial hotspots while growing the mats are inconsistent in the tensile strength test (also because of undefined directions of layers of cellulose)
• The bacterial cellulose mats showed no sign of wear on the Martindale test due to the surface being extremely smooth and not offering much possibilities for abrasion
• The Flexibility and thickness is currently to low

Reflection

The first meeting with Marc Neumann was invaluable for us to learn more about the material we try to improve. Furthermore, we learned that es expected the flexibility of the textile needs to be improved, but we were surprised by the stability at the abrasion test. This means that thick unaltered bacterial cellulose mats have quite a long longevity which is an upside because of the high price they come with. Furthermore, that bacterial cellulose mats are quite stable showed us that fashion does not have to be the only area of application and they could also serve as a form of packaging material if the necessary properties would be enhanced.

Managing Director of Cotonea: Roland Stelzer 12.09.2024

Why did we meet?

To further pinpoint the applications and a possible space in the current textile market. We wanted to talk to an expert from the textile industry. Further we wanted to talk to an industry which focuses on ethical and organic production. We hoped for insights into the current industry as well as the demands of customers. We wanted to learn what motivates customers to certain products and what the thriving factors for those decisions are. Lastly we wanted to learn what we have to do to find our place in the market and the application best suited for our product. To gain this knowledge we talked to Roland Stelzer, the managing director of the company “Cotonea”. Cotonea focuses on the beforehand mentioned ethical and organic production that is also validated by the GOTS (link meeting). Due to his position and experience in the textile industry Roland Stelzer is extremely knowledgeable in terms of market needs and customer desires.

What did we learn?

• Most consumers only care for “cheap” clothes, disregarding any production standards (this applies to the usage of toxic chemicals or GMO usage) and ethics.
• In return only about 25% of the customers demand standards like fair wages, organical products or quality.
• Our product is still only a prototype, we need to lower the production costs to be competitive with traditional companies since especially blended materials have an extremely low production spot. Furthermore we need to work on scaling up our production, especially producing big pieces of textile.
• We really need to test any properties and the ”feel” of our cloth to find the right place on the market and the perfect application for it.
• Roland Stelzer emphasised the importance of testing any mechanical and chemical property of our bacterial cellulose mats. For example he suggested that our textile should withstand 30,000 to 40,000 rounds on the Marten Dale abrasion test. Furthermore he also made clear that not only the properties of our textile are important but also the feel of it, only with knowledge about those factors we can pinpoint a marketplace to work towards.

Reflection

The meeting with Roland Stelzer left us quite in two ways as one hand we learned that in his point of view the fact that our product is based on GMO production is not nearly a hindrance as we once thought. He stated during the meeting that the people who really care about where their clothes originated from is quite small and only a subset of those are actually actively opposed. The strongest deciding factor is price by far, sadly this will pose quite a challenge for us since the LCA we conducted showed that our textile is approximately 30 times more expensive than cotton. On the other hand this could as well mean that our product appears to a smaller audience whose main focus is on sustainability and regional production and those who think price is secondary. In Addition as mentioned before we are already working on lowering the price to appear to a wider audience and therefore open more possibilities on the market for our product. Another problem we need to work on is the abrasion resistance of our textile, because especially the altered mats with xyloglucan incorporated performance laid way under the threshold Roland Stelzer mentioned (30,000 - 40,000 rounds) with the first time being about 8000 rounds and the second time with the altered mats clocking in at approximately 4000 rounds on a standard cotton mesh. Because of this we are working on standardised protocols to ensure a thicker and therefore more resilient mat for abrasion tests which we believe is necessary to increase durability (link Morone) and feasibly use our fabric for actual clothing purposes.

iGEM Team VIT India 17.09.2024

Why did we meet?

During the final weeks of the project we were still facing the challenge of implementing our dying system. While the time before the wiki freeze was coming closer, the iGEM team VIT India reached out to us. They told us about their plan to genetically modify E.coli to overexpress the genes encoding BCH and ZEP which are part of the Xanthophyll pathway. These enzymes can then be used to produce carotenoids, a group of natural pigments found in plants, algae, and photosynthetic bacteria, that are widely used in the skincare, cosmetics, and fashion industries due to their antioxidant properties, vibrant color, and health benefits. Two of these carotenoids are Violaxanthin and Zeaxanthin, which the E.coli would synthesise the enzyme to produce them out of beta-carotene in large quantities Since they want to create a more environmentally friendly, scalable, and controlled approach to the normal chemical synthesis used, and we want to create a new and environmentally friendlier way to create textiles we came together and decided that both our projects would work great together. The plan was that they would take a muslin mat and colour it with the help of their product, while we would dye our product using beta-carotene as a substitute for violaxanthin.

Reflection

After a few days both our teams fulfilled their parts of the plan and had a finished product to show. Their finished muslin cloth turned out to be an orange stained success and our BC mat also turned out great as well. We found out, that even if we just wash our bacterial cellulose mat in beta-carotene before continuing with the washing protocol, the colour is incredibly colour fast. The collaboration with iGEM VIT India was an incredible opportunity to learn and discover more about natural pigments and when the colouring step of our project could take place. Thanks to VIT India we now know, that beta-carotene is also a viable component for our CMY inspired system, as it shows as a beautiful yellow colour . We can also now say, that in application we can introduce the pigment to the already grown mat while still having an impactful result.

Emilija Suvalova 22.09.2024

Why did we meet?

Global fashion brands operate complex, far-reaching supply chains that span multiple continents. The carbon footprint from transporting raw materials, semi-processed goods, and finished products across the globe is substantial. This is why we reached out to Emilija Suvalova, who studied fashion management with extensive experience working for both global textile and leather brands. Her knowledge of the industry provided valuable insights into the status quo and how our project could impact supply chains.

What did we learn?

• Supply Chain Insights:

  Established brands, particularly in leather, have historically relied on tanneries in Southern Europe, where the leather industry is closely tied to the meat industry. From there, materials are transported to Germany for final production. While this European-based assembly line raises operational costs due to higher labour prices, it maintains consistent product quality, preserving brand reputation. Though shipping materials globally adds to the carbon footprint, maintaining these operations in Europe is seen as a worthwhile trade-off for quality assurance. Emilija found our approach of shortening the supply chain by developing textiles in a lab intriguing. She also mentioned that many companies are already researching alternatives to leather, showing potential alignment with current industry trends.

• Consumer Choices:

  Traditional leather products come in a variety of colours, with tanneries working alongside dye companies to offer a broad range of options. Emilija was particularly impressed with our modular colour system using chromoproteins, which allows us to meet consumer demand for colour variety without extending the supply chain. She saw significant potential for this approach to cater to niche markets while keeping production streamlined.

Reflection

Emilija offered valuable insight into the interconnectedness of the fashion industry, namely the symbiotic relationship between the leather and meat sectors. This is also true for cotton production; it takes combined effort of different industries (agriculture, milling and dying) to produce a piece of cloth. She also highlighted several important considerations for bringing our product to market. Beyond the requirements investigated in the property testing like tensile strength and abrasion resistance, meeting all International Organization for Standardization (ISO) standards will be crucial. This is a quality control measure, essential for all textiles on the market. Additionally, she emphasised the need for improving our yield assessed from Life Cycle Assessment (LCA), which we thus decided to further investigate in the metabolic model and inducible bacterial cellulose projects. By addressing these factors, we can make a more compelling financial case to companies looking for sustainable alternatives to the current methods.

Hochschule Niederrhein: M. Sc. Marc Neumann 23.09.2024

Why did we meet?

After gaining a lot of knowledge from the data curation from the last time we performed tests with Marc Neumann, we wanted to compare bacterial cellulose mats that were grown under the same conditions but with different % of xyloglucan in their growth media (link to experiment) to bacterial cellulose mats without, to see if it alters the properties like we expected and also to compare the data (link to experiment) to the different bacterial cellulose mats we tested beforehand. Furthermore we carried out different post treatments to see how and if they affected the mats.

What did we learn?

• washing the bacterial cellulose mats with 1% NaOH solution increases the tensile strength
• 0.25% xyloglucan concentration in the media appears to increase tensile strength but decrease flexibility
• The mats without xyloglucan had the highest flexibility
• The mats were to thin and since ripped at 4000 round of the martindale abrasion test

Reflection

results have to be taken with a grain of salt and the test should be conducted again with a longer incubation time and in more media / bigger vessels so the mats are thicker. But it appears that specific concentrations of xyloglucan indeed enhance the physical properties of bacterial cellulose although they are not the properties we were aiming for. Due to the meeting with Sebastian Cocioba in the beginning of the year we experimented with glycerol as carbon source this lead to a bacterial cellulose mat that was way more flexible to touch than the mats we produced beforehand tho to verify this we would need to conduct professional testing which is not possible because of the time constraints.

Dr. Ir. Pieter Nachtergaele 25.09.2024

Followup

Following the completion of our experiment and the collection of results, we held a follow-up meeting with Dr. Nachtergaele to discuss our experimental methods and data. He pointed out that a significant amount of residual glucose remained in the media after cultivation, suggesting that our operational parameters require refinement. He recommended strategies such as reducing the initial glucose concentration or recycling unused sugars to potentially enhance yield.

Dr. Nachtergaele emphasised the importance of considering utility costs when assessing economic viability, noting that yield alone does not provide a complete financial picture. Our goal is to streamline the supply chain by conducting dyeing and production in a single location through a co-culture of yeast and K. xylinus. Once this co-culture is successfully established, we can conduct a comprehensive financial analysis that may reveal significant advantages over traditional methods.

We also provided an update on our collaboration with iGEM Dresden, where we supplied BC mats for hydrolysis into sugar substrates. This innovative approach aims to create a closed carbon cycle, allowing us to recycle sugars back to our bacteria and optimise resource utilisation.

Finally, Dr. Nachtergaele highlighted the potential for valorising accumulated dry cell mass by exploring its use as animal feed. This strategy, also known as cascading principle, could minimise carbon waste and enhance the economic value of our overall process.

Independent Researcher Sebastian S. Cocioba 27.09.2024

Why did we meet?

We wanted to have a follow up meeting with Sebastian to further discuss with him about how our project went, which hurdles we encountered and what ressources and methods are available to us and how we can utilise the failings as a learning experience for us as well for future iGEM teams.

What did we learn?

• Do not repeat experiments that often and sequence earlier
• RBS like to form hairpins and this leading to them being cut out by e coli
• Yeast that lives in a co-culture with k. xylinus in kombucha has a natural buoyancy

Reflection

While talking to Sebastian he was proud of our achievements despite all the challenges we faced while fixing the OYC-backbone provided by iGEM (link to OYC-Backbone). In addition, he was very happy for us that we managed to achieve at least part of our goals. He provided us with a lot of feedback on how to work more efficiently and how we could deal with similar challenges in another way, We are very thankful for his support and hope to continue to keep in contact as he was instrumental in shaping the project in its current form.

Modedesign-College Duesseldorf 02.10.2024

Why did we meet?

The purpose of our second workshop with the Mode Design College (MDC) was to deepen the relationship established during our first meeting and to showcase the improvements made to our bacterial cellulose (BC) textiles. Since our last encounter, our iGEM team had made significant advancements in lab research and material development, incorporating the valuable feedback from the MDC students and textile technologists. This second session provided the ideal platform to present the enhanced versions of our KlothY textiles, which featured improvements in elasticity, softness, comfort, and resilience—qualities the MDC group had identified as areas for refinement. This time, we shifted the focus of the workshop towards a more practical, hands-on approach. Rather than a theoretical introduction to synthetic biology, we brought a wide range of pre-made BC sheets for the students and experts to work with. By offering them the opportunity to freely experiment with and creatively process these improved materials, we wanted to facilitate a deeper, more interactive exploration of how KlothY could be applied in fashion. This practical focus was crucial in allowing them to experience firsthand the enhancements we had made and to consider how the material could be integrated into real-world fashion applications.

What did we learn?

• From this second workshop, we learned that maintaining continuous engagement is key to fostering innovation. Revisiting the same group allowed us to build on the foundation laid during the first session, and this continuity opened the door to even richer feedback and creative input. The students and experts, now more familiar with synthetic biology and our bacterial cellulose material, were able to delve deeper into their creative processes. With a more refined textile in hand, they could explore new ideas, providing us with a more detailed and thoughtful range of suggestions than before. The improvements we made—enhanced elasticity, comfort, and resilience—allowed them to push the boundaries of what was possible with our material, generating new concepts and applications.

• One key lesson was the importance of iterative engagement. By returning with updated materials and inviting the students to work with them directly, we created a dynamic learning loop. Not only did this second session generate more refined feedback, but it also provided the students with the tools they needed to work more efficiently and creatively with the textile. The practical nature of this workshop gave them the freedom to explore the limits of our material and suggest further innovations based on their hands-on experience.

Reflection

This second workshop reinforced a crucial insight for synthetic biology and biotechnological projects: no matter how well-researched or well-established a project may be in the lab, maintaining an ongoing dialogue with external professionals is essential for success. Particularly when scaling up a product for real-world application, especially in the fashion industry, continuous engagement with designers and industry experts is necessary. Their input is invaluable in ensuring that a product like KlothY is not only scientifically sound but also viable in terms of usability, practicality, and design aesthetics. A key technical insight from this workshop was the use of NaOH to bleach and whiten the textile mats, which significantly facilitated the dyeing process with alcohol-based dyes. These dyes proved to be highly effective for coloring the material. However, the NaOH treatment negatively affected the sewability of the mats, making them less suitable for stitching in their dry state. Interestingly, when the NaOH-treated mats were moistened with water, their sewability improved, making them more versatile for practical applications. Another notable improvement was the use of beta-carotene-infused mats, treated with glycerol. This not only increased the material's flexibility but also provided a natural color, which was very well-received. The plasticity and moldability of the material also opened up exciting new possibilities, enabling designers to experiment with shaping and crafting new fashion projects in innovative ways. Moreover, the translucency of some of the mats added another layer of diversity to the fabric, allowing for creative exploration in design. Additionally, the ability to create specific surface patterns during the drying process, depending on the underlying surface, provided a significant advantage over conventional textiles. This workshop had a profound impact not only on us but also on the MDC students and experts. Their exposure to KlothY and the possibilities of bacterial cellulose inspired them to think more critically about sustainability and the biodegradability of the materials they use in their work. The improvements we introduced made them more receptive to the idea of integrating new, innovative, and sustainable materials into their designs. It also encouraged them to approach future fashion innovations with a more experimental and open mindset. In conclusion, experience emphasized that synthetic biology can play a pivotal role in shaping the future of fashion, but only through continuous collaboration between scientists and designers can true innovation be realized.

Conclusion

Throughout the course of our project, our approach to human practices evolved significantly. Initially, we were heavily focused on the scientific and technical aspects, consulting with researchers and experts closely tied to our university. This phase was crucial for developing a solid understanding of the core technology behind our material, KlothY. However, as the project progressed, our perspective began to shift. We started engaging with a broader community that included sustainability advocates, industry experts, and potential consumers. This allowed us to see beyond the lab and technicalities, bringing into focus the global impact our project could have, both positively and in ways we had previously overlooked. A major turning point came when we realized that some of our early assumptions and messaging were overly simplistic, and even somewhat misleading. Initially, we promoted the fast production capability of KlothY as a solution to fast fashion. However, we came to understand that fast production, while valuable for industrial applications, doesn't necessarily address the deeper, more systemic issues of fast fashion. Fast fashion isn’t just about how quickly materials can be produced; it's driven by high-paced consumerism, where low-quality garments are purchased frequently, worn briefly, and disposed of, often in landfills due to the challenges of recycling blended materials. Even though KlothY is biodegradable and locally producible, this alone doesn't make it truly sustainable. As Prof. Morone pointed out in one of our expert consultations, the most sustainable fashion choice is often to buy second-hand clothing and keep it in use for as long as possible. This made it clear to us that the sustainability of our material isn’t just about its production or disposal but also about addressing broader societal issues, such as overconsumption and waste. With this in mind, we refocused our efforts on several key areas to ensure that KlothY could be a genuinely sustainable alternative:

A. Improving Material Integrity: We worked to better understand the physical properties of our material to increase its durability and ensure a longer lifecycle, addressing one of the core issues with fast fashion - short-lived, disposable clothing.

B. Modularity and Alternatives to Blended Fabrics: We explored how to modify KlothY in a modular way, incorporating hemicellulose into our bacterial cellulose mats. This would allow us to offer a viable alternative to blended fabrics, which are notoriously difficult to recycle.

C. Resource Efficiency in Production: We also sought to reduce the resources needed to produce KlothY. By switching from a constitutive to an inducible promoter, we could better control cultivation and production, thus minimizing waste and energy use.

In addition, based on feedback from multiple experts, we began conducting a Life Cycle Assessment (LCA) to gain a deeper understanding of the environmental impact of KlothY from production to disposal. This holistic approach helped us refine our project further, ensuring that we weren’t just addressing one part of the problem but were considering the entire lifecycle of the product. Ultimately, our conclusion is that creating a truly sustainable product like KlothY requires more than just scientific innovation. It demands close interaction with a wide range of stakeholders - end consumers, fashion industry professionals, sustainability advocates, and textile experts. Only by considering the needs and behaviors of these groups can we avoid falling into the trap of greenwashing and offer a genuine alternative to conventional eco-friendly textiles in the fast-paced fashion industry. This integrated approach gives KlothY a real chance to stand out as a meaningful, sustainable solution, rather than just riding the wave of green marketing.

Value proposition

Customer Needs

Main Jobs to be Done

• Environmentally friendly clothing.
• Demand-based. No long wait times are needed.
• low price

Obstacles, challenges, frustrations (Pains)

• Conflict between wanting to stay on trend and wanting to be environmentally friendly
• common textiles using harmful chemicals
• common textiles produced far away leading to long transport ways
• uncomfortable clothing
• high price points

Benefits, Outcomes, positive results (Gains)

• environmentally friendly product
• modular product
• locally produced product

Product / Service map

Product and Services

• Produce local textiles
• Bacteria that produce bacterial cellulose naturally
• Modifiable via coculture, making a final product just as the customer needs it

Pain reliever

• Short growing time (ca. 10 days) allows for the product to be produced quickly.
• Fabric is made out of materials that are "naturally" produced and can therefore be more easily recycled than blended materials.
• The product does not need location-specific growing conditions and can be therefore produced from anywhere.
• Base product bacterial cellulose hypoallergenic. Due to modularity, many irritants can be left out to avoid skin irritation.

Gain creator

• The product does not rely on long transport ways and production steps to become wearable.
• Product is modular and properties can be changed depending on what and what concentration of inducers are given.
• The product does not use harmful chemicals in production.

The Value proposition canvas is a tool often used by Corporations to guarantee that the product aligns with the customer's values and needs. As part of our Human Practices, we would like to look at what our Project can offer and if it aligns with customer needs.

Customer Jobs - Product.

In our case, the product the customer needs are environmentally friendly clothing, that are produced exactly to the specific properties the customer needs. KlothY aims to create exactly that, by producing textiles out of Bacterial Cellulose, synthesised by K.xylinus and adding properties via a coculture.

Pain - Pain reliever

The main frustrations customers might be feeling right now is the conflict between wanting to stay on trend and still wanting to be environmentally friendly. Common textile production methods include harmful chemicals and long transport ways, especially when dealing with affordable clothing and fast fashion. A big issue customers are also facing is the texture and the feel of their clothing since uncomfortable textiles are generally not wanted by customers

Klothy is in many regards the solution to that. Since our product grows fast within ca. 10 days, and it is not bound to any specific location, long transport ways can be entirely avoided and the textile can be produced locally. Also since the product is produced in a coculture with inducible properties the feeling of our cloth as well as general properties like colour or water repellency can be modified to customer needs. The base product Bacterial Cellulose is also regarded as hypo-allergenic. Together with the modularity of our system any property that might irritate the customer can be left out. Due to the components being "naturally produced" recycling would be a lot easier since most of the current clothing produced is made out of blended materials ( natural fibres woven together with synthetic materials) and therefore a lot harder to recycle.(Niinimäki et al, 2023)

Gain - Gain Creators

As mentioned beforehand customers wish for an environmentally friendly product, that has specified properties like colour generally or water repellency in raincoats and is ideally locally produced. As outlined before our product does not rely on long transport ways, since they can be produced locally and therefore save carbon emissions compared to usual supply chains in the fashion industry. Also with the way we make our products modular, by synthesising the specific pathways in a coculture and expressing those, harmful chemicals can almost be entirely cut out.(Niinimäki et al, 2023)

As of right now, most of right now, we can not fully account for the price of the product and are still in a proof-of-concept state with our project. A future perspective for this product is not only to work on further milestones in the laboratory but also to work on price calculations and how to make this project more price effective, described further by our LCA model.

Avoid Greenwashing

One of our goals as team KlothY is to be open and transparent about our Product. Since our product is not yet ready to be commercially released and still needs further research and investigation to become so, we cannot allow companies associated with the textile industry to promote our product to greenwash. Our product is not an excuse for companies to continue environmentally harmful practices and is not an excuse for companies and legislation to continue those.

References

Niinimäki, K., Peters, G., Dahlbo, H. et al. Author Correction: The environmental price of fast fashion. Nat Rev Earth Environ 1, 278 (2020). https://doi.org/10.1038/s43017-020-0054-x