Description
The Problem
There is a pressing need for systemic change in the fashion industry. Fast fashion has made it easy for us to keep up with the latest trends, allowing for constant self-expression through clothing. However, behind the appeal of cheap and trendy fashion lies a serious environmental problem. Fast fashion enables consumers to express their individuality through constantly changing clothing, yet the cost to the environment is massive. Each year, 92 million tons of textiles end up in landfills, while the fashion industry accounts for up to 10% of global CO2 emissions (Niinimäki et al., 2020). This „buy and toss” mentality, driven by quick production cycles and low prices, creates an enormous amount of waste.
The production process itself is a significant environmental offender. The fashion industry is the second-largest consumer of water, utilising approximately 1.5 trillion litres per year and contributing around 20% of industrial water pollution from textile treatment and dyeing. Synthetic fabrics like polyester are cheap to produce but extremely harmful to the environment. Made from petroleum, polyester releases large amounts of CO2 during production and does not decompose easily, contributing to the growing problem of microplastic pollution (Niinimäki et al., 2020). Moreover, the complex global supply chains used to manufacture these clothes result in high carbon emissions. As supply chain expert Emilija Suvalova points out „The substantial carbon footprint from transporting raw materials, semi-processed goods, and finished products across the globe emphasises how the complex supply chains in fast fashion significantly contribute to environmental degradation.”
Professor Dr. Morone, a professor of Economic Policy at Sapienza University of Rome suggests that the solution lies in adopting more sustainable materials and production methods. Shortening supply chains, producing locally and using innovative technologies could drastically reduce the industry‘s impact on the environment. Professor Morone noted, „Educating consumers through tangible evidence of our product’s benefits is crucial, as many are unaware of their environmental impact demonstrating the advantages of sustainable practices over traditional methods can drive behaviour change.”
To truly reduce the fashion industry’s environmental footprint, we must rethink the way clothes are made, sold and consumed. The future of fashion needs to prioritise sustainability over speed because the planet can no longer afford the hidden costs of our fashion industry.
Our Project Description
Having this in mind, we reached out to Prof. Dr. Markus Pauly, head of Plant Cell Biology and Biotechnology Institute at Heinrich Heine University Duesseldorf, who also later became our Principal Investigator. He introduced us to Komagataeibacter xylinus (K. xylinus), a bacterial species known for its cellulose producing abilities. Bacterial cellulose (BC) pellicles hold great potential as a bio-based material with different applications. Previously, researchers used strains of Komagataeibacter to produce cellulose for industrial uses. Works of Prof. Tom Ellis from Imperial College London (Walker et al., 2024) and iGEM Vilnius 2023 and KCIS-Xiugang-Taipei 2023 are clear examples of this. Others have tried to improve the mechanical properties by infusing hemicellulose into growth media (Mikkelsen & Gidley, 2011). The Property Testing subgroup aimed to tackle a few questions: to what extent does hemicellulose such as xyloglucan improve mechanical properties? At which concentration do we see the optimal performance and does this influence the chemical composition of BC pellicles? To answer this question, we consulted experts in property testing: Dr. Amanda Staudt of DWI - Leibniz Institute for Interactive Materials. We gained insights into different types of mechanical testing and it was later conducted at Niederrhein University of Applied Sciences. We examined the change in mechanical properties, from different growth media, washing and drying procedures. We discovered that incorporating xyloglucan into the BC mats increased tensile strength at lower concentrations, though higher concentrations reduced elasticity. We also found that treating the mats with NaOH solution and drying in a cabinet at 50oC improved tensile strength. To conduct these tests on-site, we built a self-designed property testing machine, allowing us to perform mechanical tests such as tear and tensile strength tests. While most mechanical tests on BC pellicles were done at Niederrhein University of Applied Sciences, the machine provided data on cotton samples (see Property Testing). We also wanted to know the chemical properties of our mat and wanted to know if adding xyloglucan to media really had an effect. High performance anion exchange chromatography (HPAEC), experiments showed increased xylose and glucose for samples prepared with xyloglucan. We also tried to extract xyloglucan directly from tamarind seeds. However, after a long and laborious process, only a very low yield was collected.
Using synthetic biology, we wanted to produce xyloglucan in yeast in a higher amount. Contacting a researcher, PhD Ronja Ronja Catharina Immelmann we learned, which difficulties research in the area of hemicellulose production in yeast has already faced. Based on that information we worked on a pathway, to express genes involved in hemicellulose production, in Saccharomyces cerevisiae. We conducted multiple transformation evaluations and digests, to construct transcriptional units, which are part of this pathway.
Created with BioRender.com
We found out that the part BBa J435281 which we planned to use as a backbone had some errors. Sadly, we noticed that the overhangs of the OYC-eforRed-dropout plasmid backbone (BBa_J435281) do not fit with the insert overhangs for level 1 assembly.
To clarify whether the plasmid sequence was actually incorrect, or only the annotation was faulty, we transformed the parts into E. coli, miniprepped them and had the plasmid sequenced twice. (See Backbone fixing study) It appears that the uploaded sequence is correct and thus the plasmid sequence is actually incompatible with level 1 assembly, making the whole Open Yeast Collection unusable. This error appears also using the original plasmid backbone sequence deposited on the Freegenes website. We proceeded to fix the backbone by running two self-ligating Golden Gate reactions to achieve a suitable level 1 construct afterwards. As this was not foreseen before planning the lab work, at this advanced point in time in relation to the project, we decided the completion of the hemicellulose aspect of the project was not realistic and would cost too many resources, so the focus was set onto the other aspects of the project.
After discussing with Prof. Dr. Danny Ducat, we learned more about co-culture. We aimed to make stable co-culture between our engineered K. xylinus and S. cerevisiae, work by investigating their growth behaviour and optimising ratios between both of them to achieve a consistent product. (See co-culture)
In addition, we wanted to express the cellulose binding domain also used for binding the chromoproteins to our mat on the yeast cell wall directly, hypothetically increasing S. cerevisiae concentration in our mats because the engineered yeast binds to the mat directly. We hope this would potentially further optimise incorporation of hemicellulose and chromoproteins in our BC mats.(See sticky yeast)
S. cerevisiae is not only useful for hemicellulose production but also for expressing dyes to colour our fabric. In our meeting with Sebastian S. Cocioba we had the opportunity to learn about chromoproteins. When mixing them in the right ratios using inducible promoters (Park et al., 2023) we can recreate a colour spectrum similar to the one found in a printer. The colouring system used in printers is also known as the CMYK colour scheme, since it uses the colours Cyan, Magenta, Yellow and “Key” (Black). CMYK is a subtractive colour model, meaning that when none of the colours is present the displayed colour is white while combining every colour results in a black colour being displayed. Since we also learned in the meeting that chromoproteins denature shortly after being expressed, we decided that pigments would be a much more stable and viable colouring agent for the future. We believe that future iGEM teams can take on the challenge of finding the perfect pigments and engineering a suitable pathway for them.
Created with BioRender.com
But how sustainable is our process? From our conversation with Global Organic Textile Standard, we found that sustainability models, such as life cycle assessment (LCA) can help in making such a judgement. We talked to an LCA expert, Dr. Pieter Nachtergaele, and conducted LCA experiments to determine the yield. However, the predicted yield was too poor to make our process economically viable. One way to overcome this is by lowering the substrate cost. We got in touch with iGEM Dresden, whose project focused on hydrolysing clothes to produce sugar substrates. If this sugar is fed to K. xylinus to produce new clothes, we have achieved a closed carbon cycle - solving both economic and sustainability issues. Another way is to make a metabolic model, which helps us understand different flux modes in K. xylinus. From this, different strategies to maximise cellulose flux can be devised. We were able to identify candidate reactions that can be knocked out and knocked down to optimise cellulose flux close to its theoretical maximum.
One of the major difficulties in researching and working with K. xylinus currently is the constitutive bacterial cellulose production which while quite performant also makes it difficult to properly cultivate in higher cell concentration, as K. xylinus tends to undergo spontaneous non-cellulose mutation during shaking cultivation (El-Gendi et al. 2022) reducing the viability of the culture to produce consistent mats unless expensive cellulase is added. In addition, we were also worried that to reach better incorporation of hemicellulose and dye the bacterial cellulose needs to be regulated. This is why we were interested in an inducible system and successfully generated a K. xylinus a knockout strain and an inducible strain. With that we could eventually reduce the need for cellulase necessary to increase cell count and optimise carbon intake by targeting knockouts or overexpression for different genes and or clone an inducible promoter in front of the gene we want to up or down regulate, giving us even more control over the bacterial cellulose synthesis. Through this we hope to push the scale in our favour to achieve a truly sustainable product.
Detailed Property Testing Description
Abstract
The goal and purpose of the property testing subgroup was to get information on how our produced textile behaves compared to commonly used textiles like cotton and polyester. For this, we tested the mechanical and chemical properties of the Bacterial Cellulose (BC) mats we created under varying conditions. In the context of composition analysis using HPAEC, we could prove that xyloglucan was implemented in the Bacterial Cellulose mat when growing it in media with 0,5% (w/v) of xyloglucan (XG) or washing it with a 0,5% (w/v) XG solution. There, the effect of growing the mat with XG was greater than washing it with a XG solution. We further hypothesized that the intercalation of hemicellulose in the BC mat would increase flexibility and tensile strength. This goal was partly achieved, as we found that a xyloglucan concentration of 0,25% increases the tensile strength, but reduces the elasticity. For higher concentrations, we found that both tensile strength and elasticity are reduced. Furthermore, we tested the effects of different washing and drying methods on the mats’ properties and found that washing the mat with a 1% (w/v) NaOH solution and drying it in the drying cabinet increases the tensile strength. In addition, we built a simplified property testing machine that can perform tear tests, as well as tensile tests, and may help future teams that need such tests but lack funding and/or contacts with professional machines.
How property testing started:
"There was an idea, [...]. The idea was to [...] [evaluate and compare the textiles we grow to commonly used textiles like cotton and polyester] to see if they could become something more. See if [...] [we could improve them to be more durable and eco-friendly]"1. For this, it was crucial to know how durable our textile was in the first place, meaning we first had to agree on what properties we would deem essential to evaluate these in the form of tests. Searching through literature describing the art of property testing on textiles 2, we first collected properties that we regarded as crucial. These included mechanical properties 3 like abrasion, tensile strength, tear strength, axial torsion, and puncture resistance, chemical properties like composition, washability, and flammability 4, and also properties regarding the textile's general performance 5 like water repellency, breathability, and UV absorption. To evaluate our data we wanted to apply mathematical modeling. The goal was to predict the property outcomes from cultivation variables and find the "sweet spot" in terms of cultivation parameters and post-treatment. In terms of mathematical models, we wanted to make use of linear regression. To reduce the tested parameters for easier data comparison, we later reduced the tested properties to abrasion, tensile strength, tear strength, and chemical composition. We then researched the necessary tests. We chose the Martindale method 6 (ISO 12945-2:2020) to test the abrasion of our textiles and the Trouser Tear (Single Rip) Test 7 similar to the one described in ISO 34-1 8 for testing the tear strength. For tensile strength, we tried to replicate tensile tests as described by Instron 9, for example, as it was hard to find a specific method to go by in this area. Some adjustments had to be made in mechanical testing as big pieces of our cloth were harder to make due to limited space in the incubator. That's why we, for example, chose the single rip method 7 (ISO 34-1 8), which is usually used for plastics and rubber to address the tear strength instead of the Tongue Tear (Double Rip) Test 7 (ISO 13937-2 10), which is the one that is usually used for textiles but needs larger samples. For composition analysis, we chose to do high-performance anion-exchange chromatography (HPAEC). The problem that arose then was that we didn't have the machines necessary for the mechanical testing at our university. That's why we searched for institutes at other universities with specialized machines. Searching for an institute with the necessary machinery, we contacted Amanda Staudt from the DWI - Leibniz Institute for Interactive Materials in Aachen, who helped us work out our ideas for mechanical and chemical property testing. She also advised us on what to look for in cultivation and mathematical modelling. From her, we also got the necessary measurements and the number of replicates for our mechanical tests (Abrasion, tensile strength, and tear strength). Then the Institute of Textile Technology at RWTH Aachen, which we contacted shortly after, told us that we could test to chemically treat our BC mats to increase their hydrophobicity (something that would be useful if we for example wanted to make outdoor clothing with our mats). They also gave us advice on testing the purity of our products, as their research on their LignoTex 11 project indicated, that removing impurities like lignin and hemicellulose results in stronger fibres (something that was in some sense contrary to our project of improving the flexibility of our BC mats by adding Hemicellulose). Furthermore, they advised us to try solubilising our Bacterial Cellulose to convert it into fibres and yarns as they could then be interwoven with other fabrics to get the best of both worlds. The problem there though, as we later found out, was that the hemicellulose in our textile would degrade in the process, which was why we scrapped that idea. While preparing our first round of BC mats to test in Aachen, we also built our own hardware. This act was inspired by the advice of one of our PIs, Prof. Guido Grossmann, who suggested that we could perform simplified versions of the mechanical property tests ourselves for easier and faster data access. What his suggestion led to, was the creation of our self-built property testing machine.
What we did:
Once the first small Bacterial Cellulose mats had been grown in the lab, our first goal was to make bigger versions that could be used for mechanical testing. With this, we first wanted to explore their potential for the future of our project and get used to handling them right. This way, we grew our first round of mats in a baking dish and a 15x15 cm glass container at room temperature due to limited incubator space. In the process of growing the mats, we could already see that they grew better on YPD Media than on DSMZ Media. We then tested how they would react to autoclaving and washing as we feared that they would not survive the harsh conditions of the autoclave. The extraction of the mat grown on YPD before autoclaving can be seen in picture 1.
Picture 1: Bacterial Cellulose mat, grown in YPD media for six days, being extracted.
The autoclaving process did not destroy the mats. The only visible change was that the edges appeared to be a little bit dried out. After later also extracting, washing, and drying the mat grown on DSMZ, we could see that it was a lot thinner and less robust than the one grown on YPD, as it already ripped apart in the cleaning process. On the flip side though it wasn’t stained by the media like the one grown on YPD and was more transparent (Picture 2).
Picture 2: Bacterial Cellulose mat, grown in DSMZ media (left) and YPD media (right), after drying. The mats were washed repeatedly with cold deionized water.
In the meantime, a big Bacterial Cellulose mat was grown for an extended time to see if it would change its properties. The result was that the mat got thicker and stiffer with the extended cultivation time.
The next step was to find out how the cultivation parameters could be perfected for the best properties of our mats. Following a paper [Mikkelsen et al., 2011 12] on growing Bacterial Cellulose mats with hemicellulose, we grew these mats on HS media. The media was then modified by switching from glucose to glycerol, as we wanted our carbon source to be more sustainable. If glycerol worked, we could grow our mats on waste products like crude glycerol in the future. To find the right amounts, we grew the mats at glycerol concentrations reaching from 1% to 4%. The other half of the experiment was to test the effects of different concentrations of the Hemicellulose xyloglucan on the mechanical properties (in this case abrasion & tensile strength), with concentrations ranging from 0,1% to 0,5%. What we also wanted to find out with composition analysis via HPAEC, was, if the Hemicellulose would be incorporated in the BC mat and if the amount would increase with increasing concentrations in the media. Both plans for this experiment were combined by either making the glycerol or the xyloglucan concentration the variable with the other variable being at the maximum concentration for the experiment. While cleaning and drying the mats, we could already make out differences in the mats' properties like thickness and how easily they would break down, which was what happened to some of the mats to different degrees, making 3 of them unextractable. In general, samples grown in lower xyloglucan concentrations (0.1~0.3%) exhibited thinner and flimsy attributes than those grown in higher xyloglucan concentrations (0.4, 0.5%).
The problem that then faced us, was that DWI cut our cooperation, meaning that we couldn’t do mechanical tests on our tests as planned. As the self-built property testing machine also wasn’t finished at that point, we again searched and reached out to other institutes to perform our mechanical tests. After some time we got in contact with Marc Neumann at Hochschule Niederrhein (HSN), who generously offered us to do these tests at their institute. We were also offered to perform our mechanical test at Hochschule Düsseldorf (HSD), which we didn't get to though, due to scheduling problems. The first time we went to HSN, we performed Martindale Abrasion tests and Tensile Strength tests on the mats from the experiment with differing amounts of glycerol and xyloglucan as well as on mats from other experiments (Pic. 3). Additionally, we tested a cotton cloth we chose as our standard of comparison and pieces of Aramid/Kevlar.
Picture 3: Tensile strength test (left) and Martindale abrasion test (middle) performed at HSN, and tear test performed on self-built property testing machine (right).
The remnants of the tested BC mats were then sent to iGEM Dresden 13 as they work on textile degradation and could find out if the textile can be recycled in a meaningful way. Something that was also interesting to look at, was a Bacterial Cellulose mat, which had been unknowingly contaminated with a fungus that was washed and dried for the means of testing possible property changes. Looking into the data, we couldn't draw any definite conclusions, as the standard deviation was too big, or in the case of cotton not calculable (Fig. 1).
Figure 1: Tensile test data from the first round of tests at HSN. The graph shows the standardforce by thickness in N/mm (blue) and the elongation by thickness in %/mm (orange) for different textiles. Those textiles include 100% Cotton, Aramid, and BC Mat grown on YPD media, YPS media, and YPD media contaminated with fungus. The standard deviation is shown in red/black. For cotton, no standard deviation could be calculated, as there was only one sample.
Before the tests at HSN, we performed a first round of HPAEC, to find out how the concentrations of glycerol and xyloglucan affected the composition of our mat, and to find out if xyloglucan was integrated at all. After the HPAEC we could see from the peak in Xylose that xyloglucan was incorporated, but Ribose which we used as our standard, was also found in the BC mat, meaning that we would have to change our standard. The problem with the standard also made a statistical analysis of the data impossible, as we couldn't reliably calculate the amounts of our sugars. The problem led to us growing new mats for two more rounds of HPAEC. This time we grew mats with less variation, as the powdered xyloglucan we used is quite expensive. The standard was changed to Fucose, as we knew from the previous round, that it wasn't present in the BC mat. The results are summarized in Figures 2 and 3.
Figure 2: Sugar composition of Bacterial Cellulose mats, grown/washed with different amounts of xyloglucan (XG). The concentrations have been measured via HPAEC with Fucose as standard.
Figure 3: Glucose and Xylose concentrations of Bacterial Cellulose mats, grown/washed with different amounts of xyloglucan (XG). The concentrations have been measured via HPAEC with fucose as standard.
The results of the HPAEC were as follows. No Xylose could be found in the BC mat grown without xyloglucan (0% XG), while there was an intermediate amount in the mat washed with 0,5% (w/v) XG solution (0% XG washed in 0,5% XG) and a relatively high amount in the mat grown with 0,5% xyloglucan (0,5% XG), which means, that xyloglucan was incorporated in the Bacterial Cellulose mat (Fig. 3), as the Xylose could only come from the xyloglucan and is therefore not present in mats grown/washed without xyloglucan. We can additionally say, that the effect of growing the mat with XG is greater than washing with XG solution. An increase in other sugars like glucose, galactose, mannan, ribose, as well as galacturonic acid could also be documented for the mats grown with 0,5% (w/v) XG (Fig. 2).
Something that we also successfully tested was washing our BC mat with a 1% (w/v) NaOH solution as suggested by literature sources [El-Gendi et al., 2022 14]. There we found that the NaOH helped in decoloring the mat, which led us to apply the method for other experiments that followed. Additionally, we performed a test intending to find the perfect amount of media for cultivation. The results from this were that mats seemed to grow best with the container filled to 33% with YPD media, with the rest being air.
The second time we went to HSN, we prepared BC mats, grown with different amounts of xyloglucan, to evaluate possible property changes (Figure 4). In addition, we tested the effects of different washing and drying methods on different textiles (Fig. 5 & 6).
Figure 4: Effects of different concentrations of xyloglucan (XG) on the normalized tensile strength (tensile strength by thickness) and elongation (elongation by thickness -> elasticity) of Bacterial Cellulose mats.
Figure 5: Effects of different drying conditions on the normalized tensile strength (tensile strength by thickness) and elongation (elongation by thickness -> elasticity) of Bacterial Cellulose mats.
Figure 6: Effects of washing Bacterial Cellulose (BC) mat with 1% (w/v) NaOH solution. The graph shows the normalized tensile strength (tensile strength by thickness) and elongation (elongation by thickness -> elasticity) of BC mats, grown with different concentrations of xyloglucan (XG).
Looking at the data from HSN round 2, it shows that a 0,25% (w/v) xyloglucan concentration increases the tensile strength of the BC mat, while higher concentrations decrease it (Fig. 4). Meanwhile, the elongation, which directly correlates with the elasticity, seems to decrease with the addition of xyloglucan to the BC mat. In terms of drying conditions, drying the BC mat in a drying cabinet seems to be the best method as it increases tensile strength, as well as elasticity compared to parchment drying (Fig. 5). When it comes to washing methods, additionally washing the BC mat with a 1% (w/v) NaOH solution shows to be superior to only washing mats with cold ddH2O, as it increases tensile strength as well as elasticity (Fig. 6).
Property testing machine:
The creation of the first version of our self-built property testing machine started by making a frame out of upcycled wooden planks left from construction work. We tried to replicate Universal Testing machines like this one by Nextgen [^35]. The clamps of the machine were improvised with modular metal construction materials like were sold by Eitech 15 (Pic. 4).
Picture 4: Wooden property testing machine prototype, also nicknamed “DER GERÄT”. The version in the picture has been modified, to be used as an interactive game to present the basic principles of tear testing
When we then presented the machine to our PI, Prof. Markus Pauly, who advised us that if we wanted to use the machine in the lab, we would have to either paint to machine or switch to another material as wood is porous, making disinfection a problem. This inspired us to build a new and improved property testing machine out of galvanized sheet metal, again upcycled from leftover construction material. We then added a programmable LEGO EV3 motor to this machine to apply force to our samples in a tear test. In the first tests with the motor, we ran into the problem that the string we used couldn’t be wound up properly (Pic. 5). After doing some research, we found that iGEM Team GreatBay_SZ 2019 had designed a spinning mechanism 16 that solved our problem after being adapted to our system. After solving this problem, the motor was integrated into the machine to do further tests, to also measure the maximum force output of the motor. There we only reached about 6 N which was barely enough to rip apart cardboard. This led us to redesign the mechanism, this time integrating a double motor system with stronger motors (Pic. 5). After writing the necessary code, to get the motors to spin in the right direction, the first tests with this system could be performed. It turned out that with the changes made, the force output got beyond what we could measure with our 30 N Newton meter.
Picture 5: Evolution of the LEGO winding mechanism of the property testing machine. A graphic of the iGEM engineering cycle was added digitally.
The next change that was made was to screw legs onto the construction so that when it was flipped upside down a simplified tensile test could be performed (Pic. 6). The test was modelled after a simplified tensile test described by Michigan Tech 17. To make the machine flippable, the LEGO motor was integrated in a way, that made it possible to easily take it off the machine.
Picture 6: Property testing machine mark 2 (“DAS MASCHIN”) in tear test configuration (left) and tensile strength configuration (right).
The machine was then used to get tear strength data on our 100% cotton cloth which we wanted to use as our standard of comparison (Fig. 7). This worked relatively well in weft direction, but failed in warp direction, as the clamps and Newton meter were not strong enough to hold the textile / measure the force at a point, where the textile would tear. Even though the standard deviation for the data in weft direction was relatively low, conclusions on how well the machine works compared to professional Universal Testing machines 18, as a comparative test with such a machine and with the same cotton cloth was not possible, due to organizational constraints.
Later on, the machine was also used for doing the same in the context of a tensile test. Throughout these experiments, we realized that many improvements could be made for more reliable tests. These include stronger Newton meters or, as we discussed with a member of iGEM Münster, knowledgeable in the field, we could switch to an Arduino or Raspberry Pi based system, implementing a stepper motor, giving us a digital readout in terms of the voltage change of the motor over time. With this data, we could then properly calculate the applied force at a given time.
Figure 7: Tear strength of a 100% cotton cloth in different directions (weft & warp) measured with the property testing machine “DAS MASCHIN”.
A further modification that was made on the first property testing machine was to repurpose it, creating an interactive game teaching others about the basic principles of our tests. This was already used at the sustainability day at our University, as well as at the JuniorJam in Münster.
How/What to improve in the future (Advice for future teams)
On the textile:
Testing the possibilities of reinforced textiles by letting the BC mat grow into a net. Further, explore the effects of washing BC mats with NaOH solution at different concentrations as it possibly further improves properties
On the machine:
stepper motor and Arduino or Raspberry Pi based system for digital force readout (calculating force from the Voltage needed to move the motor at a specific speed) -> more precise than Newtonmeter Stronger building material (e.g. galvanized square steel) Stronger clamps that are easier to close and open with rubber-lined teeth
References
Future Applications
In our iGEM project, we developed many tools that are necessary to push forward innovation in using bacterial cellulose as a material alternative to those that rely on conventional synthetic-dependent processes. Ranging from the hardware that can be used to conduct property tests on-site, sticky yeast for co-culture, colour software to predict the colour of the textile, and genetic toolkits to implement genetic modifications: future iGEM teams can try to implement different aspects of our project that were unfinished. Namely, knocking out those candidate genes and using our inducible system to validate metabolic model prediction and validate for flux to ensure a comparable cellulose yield in engineered K. xylinus strain.
References
Niinimäki, K., Peters, G., Dahlbo, H., Perry, P., Rissanen, T., & Gwilt, A. (2020). The environmental price of fast fashion. Nature Reviews Earth & Environment, 1(4), 189–200. https://doi.org/10.1038/s43017-020-0039-9
Walker, K. T., Li, I. S., Keane, J., Goosens, V. J., Song, W., Lee, K.-Y., & Ellis, T. (2024). Self-pigmenting textiles grown from cellulose-producing bacteria with engineered tyrosinase expression. Nature Biotechnology. https://doi.org/10.1038/s41587-024-02194-3
Mikkelsen, D., & Gidley, M. J. (2011). Formation of Cellulose-Based Composites with Hemicelluloses and Pectins Using Gluconacetobacter Fermentation. In Z. A. Popper (Ed.), The Plant Cell Wall: Methods and Protocols (pp. 197–208). Humana Press. https://doi.org/10.1007/978-1-61779-008-9_14
Park, J. H., Bassalo, M. C., Lin, G.-M., Chen, Y., Doosthosseini, H., Schmitz, J., Roubos, J. A., & Voigt, C. A. (2023). Design of Four Small-Molecule-Inducible Systems in the Yeast Chromosome, Applied to Optimize Terpene Biosynthesis. ACS Synthetic Biology, 12(4), 1119–1132. https://doi.org/10.1021/acssynbio.2c00607
El-Gendi, H., Taha, T.H., Ray, J.B. et al. Recent advances in bacterial cellulose: a low-cost effective production media, optimization strategies and applications. Cellulose 29, 7495–7533 (2022). https://doi.org/10.1007/s10570-022-04697-1
Footnotes
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https://www.instron.com/en-us/resources/test-types/tensile-test, last opened 09/26/2024, 21:45 ↩
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https://www.biosc.de/lignoTex, last opened 09/26/2024, 21:45 ↩
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Mikkelsen, D. and Gidley, M. J. (2011) Formation of Cellulose-Based Composites with Hemicelluloses and Pectins Using Gluconacetobacter Fermentation, The Plant Cell Wall: Methods and Protocols 14, p. 202. ↩
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https://2024.igem.wiki/tu-dresden/, last opened 09/26/2024, 21:45 ↩
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El-Gendi, H., Taha, T.H., Ray, J.B. et al. Recent advances in bacterial cellulose: a low-cost effective production media, optimization strategies and applications. Cellulose 29, 7495–7533 (2022). https://doi.org/10.1007/s10570-022-04697-1 ↩
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https://www.eitech.de/, last opened 09/26/2024, 21:45 ↩
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https://2019.igem.org/Team:GreatBay_SZ/Hardware, last opened 09/26/2024, 21:45 ↩
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https://www.mtu.edu/materials/k12/experiments/tensile/#:~:text=The%20basic%20idea%20of%20a,the%20other%20end%20is%20fixed, last opened 09/26/2024, 21:45 ↩
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https://www.nextgentest.com/metal-testing-equipment/ universal-testing-machines/, last opened 09/26/2024, 21:45 ↩