Team:Toulouse INSA-UPS/Human Practices
Human Practices
Welcome to our Human Practices page. Here you can find an Overview of our Human Practices effort and the Integrated Human Practices part of our project.
Overview
From the beginning, we wanted to build a project which is responsible and good for the world. This has been carried out through three main axis: integrated human practices, entrepreneurship and Education.
Integrated Human Practices
We were not expert in anything, so we had to use the experience of our supervisors and find other experts to help us (professors at INSA, people working in companies specialized in fragrance production, associations, etc).
These efforts are described in our Integrated Human Practices page.
Entrepreneurship
Thanks to these people, especially Robertet that provided us with market figures, we were able to build a solid implementation of our project, allowing to predict its consequences in the real life, and hence, proving the usefulness and goodness that our project could bring.
This work is presented in our Entrepreneurship pages.
Education and Communication
The Covid19 pandemic has confronted us with the importance of communication and explanations about biotechnologies, especially towards the general public and people whose condition is a barrier to access science knowledge. Biotechnologies are more and more used to achieve sustainable development but this requires acceptation by the general public. In this scope, we have created and used a pedagogic game to explain our subject, the world of perfume and synthetic biology in many events.
All these activities are described in our Education and Communication page.
Integrated Human Practices
We chose and engaged in a project we were not experts in, simply students with the will to create a feasible and coherent project. To do so, we contacted specialists, stakeholders and potential users at each step of our project in order to get constructive feedback and crucial advice. Integration of all these informations was the indubious key of our project success.
Here, we present the people, companies and organizations whose information shaped our project, from the first choices, its design, the final production, to its application in the real world. Thus, this page is divided into these four sections. Each section contains several steps of our project and how these steps were influenced by the advice we were given. The final synthesis exposes how Integrated Human Practices were the backbone of this project. Abstract of each interview is presented for a rapid glance. Our notes from each interview are also accessible in drop-down panes and of interest to any future iGEM team that wants to engage in the development of other synthetic perfumes.
Choices
Our initial idea was to create a yeast strain to produce a whole modular perfume, with several enzymes under several inducible promoters. We discussed this idea with the perfumer L’Occitane and Yannick Malbert, CTO from Sweetech. They suggested us to tune our project towards coherent groups of fragrance molecules (called “accords”) instead of whole perfume. Most importantly, they validated the huge potential of the system but warned us about the need to choose the target smell well to ensure the project's interest. This is how we decided to focus on the violet accord.
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L’Occitane, international company
L’Occitane is an international company specialized in the production of cosmetic products. They proposed to focus on fragrance accords, with simpler mixes of molecules (5-7 molecules) than in a whole perfume (30-200 molecules). They also hinted at the very appealing possibilities that our project offers to produce rare compounds, or those which provenance is complex or even forbidden by IFRA (International Fragrance Association).
Subject : Perfume industry and industrial demand
- Overall, they did not consider realistic the idea of a yeast producing by itself a whole perfume. Among interesting accords to recreate, they pinpointed mute flowers (lily of the valley, violet), foams, marine, musky, or solar scents (if we could develop molecules with these scents it really meets a demand from the current industry). Some accords are very expensive such as ambroxide or burnt odor (birch, cade and styrax). They were not interested in products already easily found in the food industry, such as orange aroma
- It is interesting to bring the “natural” side with the yeast. Consumers are turning more and more to natural fragrances. Our project will bring this “naturality” which currently lacks too many perfume specialities. This way, it would be possible to obtain the Cosmos certification for biological and/or natural products.
- About sustainability, any solution to replace hexane-based extraction is relevant for them. They also liked very much the idea to use green biomass as a substrate for the growth of our microorganisms.
- They raised the problematic of allergenic compounds in some perfumes. Being able to modify the molecule to get rid of the allergen could be of high interest, even if perhaps complex and risky toward the conservation of the fragrance properties.
- To the best of their knowledge, there is no similar subject such as Elixio in current development. They also inquired about intellectual property in the iGEM context and agreed to be contacted again.
Yannick Malbert, from Naturamole
We met Dr. Malber who worked for several years at Naturamole, a company specialized in the development, the production and the marketing of natural molecules for the flavors, fragrances and cosmetics markets. We explained to him our refined idea to create a sustainable violet accord. He confirmed to us that the idea has a real potential, more than the creation of a full perfume. He saw as an added-value the fact that biotechnology sourced molecules can also be considered as natural molecules. For him, this is a big advantage as more and more consumers want to have access to natural molecules and value eco-friendly production.
Subject : Perfume industry and industrial demand
- Perfumers use pure molecules that they then mix. Creating an accord is much more interesting than a perfume (a perfume is composed of a hundred of molecules). Biotech market is trendy and naturalness is sought after by consumers. It is however different for the perfume industry where packaging and branding are very important for the consumer
- Many projects are interested in the production of pure molecules. Our project is thus very different and has potential.
- How can we extract our molecules from the complex medium? Aromatic compounds are volatiles. Thus, a liquid-liquid extraction should work but we will have to use solvents. Another solution is fractional distillation but we could lose our mix with this method. For essential oils, steam drive is used.
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Naturalness is submitted to drastic characteristics :
- Natural substrates are used.
- No solvents nor chemical catalysts are used in the process.
- The produced molecules exist in the wild
- Our target molecules have antibacterial characteristics. We should perform toxicity tests on our yeasts and then define the lethal concentration. It also applies to the intermediate molecules. In general, the final enzymes in the pathway are more active than those at the beginning. This effect prevents the accumulation of toxic intermediates.
- On which molecules should we focus on? Dr. Malbert advised us to choose two flowers, for example violet and lily. Some compounds are microdosed and are very important in the smell of the flower. We have to identify these compounds. If we cannot produce these molecules, we can say that we want to produce a simple base that can be complexified if needed.
Design
The road from the initial concept (using synthetic biology to produce a violet accord) towards the complete design was a difficult one, with many challenges to face. Identification of the molecules and best enzymes was rather straightforward from the literature. However, many critical points have to be well thought, such as enzyme expression and organization (fusion or not for example), suited microorganisms for optimal expression and how to modify them, and molecules extraction. Luckily, we had many experts close to us in Toulouse Biotechnology Institute to answer our numerous questions. With their help, we built a strong project design, feasible with reduced risk for the six of us over the summer. Their interviews are summed up below.
Modelling discussion are described elsewhere since Pierre Millard, our expert, was part of the instructor team from the very beginning of the iGEM Toulouse 2021 adventure (see Modeling)
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Philippe Urban
Philippe Urban is the head of a team working on terpenes at TBI. He was really enthusiastic about our project. For him it was important to mimic the real smell of the flowers. As one odor is not due to a single molecule, but more to a complex mix of molecules, he encouraged us to produce several molecules to mimic a scent. Philippe also gave us the idea to drive the production of each molecule by an inducible promoter to fine-tune the proportion of each molecules in the mix.
Subject : Design of the yeast
- In the 1920's, to test the opening of an airline between Paris and Toulouse, a bouquet of freshly picked violets was delivered on the first flight. This is an anecdote we could use
- The subject seems very original and funny.
- Will the natural smell of the yeast be a problem? Should we try to eliminate it?
- Yeast seems to be a good organism for our terpenes production.
- An odor is not due to a single molecule. For example vanillin is the molecule mainly responsible for the smell of vanilla. However, when you smell vanillin and a vanilla bean, you can directly tell the difference because vanillin alone is not sufficient to reflect the complexity of the vanilla smell. So it can be interesting to focus on our flower and to produce several molecules responsible for the scent of this flower.
- A regulatory part could be beneficial to the project. We could play with the induction of our molecules to get closer to the real perfume of the flowers by fine-tuning the proportions of each molecule.
Gilles Truan
We discussed with Gilles Truan, head of the TBI, about the inducible promoters suitable for our system. Chemically inducible systems seemed safer for the project. As a yeast expert, he was the one to take advice from about the design of our yeast strain, notably the optimal loci to integrate DNA into the yeast genome. He explained to us that it would be better to multiply integration loci, because the length of integration is limited in a locus.
We also talked about the production of our aldehydes, and were warned that they could be toxic for yeasts. He approved our idea to produce the aldehydes in cyanobacteria and to make a consortium between these two organisms.
Subject : Yeast and high producing lycopene strain
- Modulation is very complicated, so the idea of producing different accords in one strain seems complicated. There are not enough different inducible systems in yeast. In addition, optogenetic regulation is complex and does not always work. With chemically inducible systems, it should be easier to control the expression, for example with PHO, MET, GAL (with this one, the yeast has to change its carbon source so there should be no glucose left in the medium), or TET ON/OFF. We have to think that once the inducer is introduced in the medium, it is impossible to remove it. We could also try to induce some genes according to cellular processes such as cell density.
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Gene integration: integrating 5 inserts should not be complicated,
maybe we could use Crispr. If we want to make a fusion protein it is
possible
but if it does not work, we could lose time.
- Where should we integrate our genes? There are neutral zones in terms of interference, with low transcriptional activity. Integrations can be safely done there
- Integration length? There is no known limit for the number of integrations in the yeast genome. This could be a problem for E. coli manipulations because long strands of DNA are fragile. We want to integrate 5 different genes, we could do it 3 at once and then 2, or 2 at once then 2 and then 1
- Analytical part: the carotene color is visible by eye. With HPLC, all the intermediates are detectable because they are all of a different color. Separation protocols are widely documented. Analyzing ionones with HPLC is however impossible, because these compounds are very volatile. We must also recover ionone from the medium without losing too much of it during the culture.
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Aldehyde production:
- Yeast: the yeast membrane is composed of linolenic/linoleic acid. When the yeast produces carotene in large amounts, it is transported into lipid droplets so there is production of linolenic/linoleic acid. If we decide to produce aldehydes with S. cerevisiae, we must introduce two more enzymes into the yeast genome. Moreover, aldehydes are very reactive and therefore often toxic in vivo..
- Cyanobacteria: it is possible to produce aldehydes with cyanobacteria, grown in a consortium with yeast. We must however keep in mind that the cyanobacteria division would be the limiting step (far slower than yeast).
- Recovery processes: it is possible to recover the molecules directly in the organic phase. The culture can be done in 10% dodecane. But it is not easy to recover dodecane and then we must extract odorant molecules from this solvent.
Jean-Marie François
We contacted Pr. Jean-Marie François from TBI to talk about the design of our engineered yeast, Saccharomyces cerevisiae . First we discussed the inducible part in order to produce our molecules. With his advice, we identified 4 induction systems for the production of our 4 terpenes. The induction molecules selected for the project were galactose, doxycycline, copper and estradiol.
Jean-Marie also gave us tricks in order to integrate our constructions into the yeast. He told us to perform transformations with linear fragments with long enough homologous flanking ends.
Subject : Yeast and biochemical pathways
- Low metabolic flow in the mevalonate pathway : the pathway should be promoted with a channelling.
- Regulation is a difficult part. In addition to the promoter, another key to a tight regulation could be to adjust enzyme kinetic parameters. Sugar-inducible promoters may not be the best solution because of metabolic switches that could occur with the addition of another carbon source. During co-culture, galactose-inducible promoters could however be a great option. Indeed, cyanobacteria would produce a slight amount of sucrose (=> glucose), so the system should be able to sense and response to galactose. Galactose-inducible promoters have been extensively studied in yeast. For the theoretical part (without co-culture), we could grow our yeast on raffinose and test the different galactose promoters. Doxycycline, copper and B-estradiol promoters are otherwise great choices of regulatory systems
- The transformation can be done with linear fragments with long homology regions (100-500bp). Classical recombination methods work very well in yeast. We need to identify gene targets for a possible integration, for example we could delete a gene from another metabolic pathway not essential (pyruvate dehydrogenase or alcohol dehydrogenase) with the transformation.
Sara Castaño Cerezo
Sara provided us with a yeast strain producing a high amount of lycopene, and we had the opportunity to discuss with her about our design. We talked about the cloning methods and optimal integration loci to be used. She recommended us a key paper describing useful plasmids and associated integrative loci. She told us that the yeast could grow with sucrose, which was perfectly in line with our sucrose producing cyanobacteria and yeast consortium.
Subject : All about S. cerevisiae
- Sara provided us with a high lycopene producing strain derived from the CNPK yeast strain. These strains are excellent and robust terpenes producers. They also can use sucrose as a substrate.
- There is not so much production of GPP so we could encounter problems with the production of linalool and other GPP derivatives. In order to produce more GPP, the enzyme GPPS should be mutated.
- The HO locus is already occupied in the lycopene strain. We have to find another site to integrate our inserts but it should not be that difficult.
- She proposed to use bidirectional promoters to limit the number of promoters to define and putative interferences.
- A fusion enzyme between LcyE and ofCCD1 was developed in E. coli. A problem in the yeast could be a low expression. Regarding the size of the linker in E. coli, she could not guarantee it would be suitable for S. cerevisiae. The only way to know is to try. She did not know if the other fusion (CCD1 + crtY) would work. Once again, tests must be carried out. The more we test, the more we have a chance to obtain a functional construction.
- She did not like the TET promoter. The expression with this promoter is not high and the doxycycline is not very stable in the yeast (the results are not reproducible). She thinks it should help to add a copy of TETr in our construction
- What type of cloning method should be used ? She uses the Gibson method but digestion-ligation should also work. The Gibson method is however faster because there are fewer steps. Cloning with the pUC19 plasmid is a good idea, she uses this technique too. She finds all of her plasmids and markers in papers from Irina Borodina (easyclone).
Denis Jallet
As we decided to use a consortium between a yeast and a cyanobacterium, we needed the opinion and advice of cyanobacteria experienced ally, and were lucky enough to meet Denis, a researcher at TBI. He warned us about the long growth time of cyanobacteria, a risk for us since we had limited time over the summer to perform our experiments. He advised us to start growth tests in parallel with the genetic engineering of the cyanobacterium. For him the strain we identified, UTEX 2973, seems to be very interesting with its fast growth rate. He listed us the required equipments to cultivate the cyanobacteria, especially to provide them with sufficient lighting. Eventually, we convinced him to join the team of our instructors.
Subject : Cyanobacteria
More details
- We must carefully choose the strain we want to work with; each cyanobacterium has its own specificities !
- We have to be careful about time management since slow growth often limits experiments, and that could be the case with cyanobacteria. Usually, two months is the minimum required between transformation and observation of results. We should expect one month to grow the wild-type on petri dishes
- In order to save time, the cloning part of the cyanobacteria should be done in parallel with growth tests.
- We introduced to him the cyanobacterial strain UTEX 2973. For him the strain looks very interesting as it seems to grow faster than most cyanobacteria.
- UTEX 2973 grows faster but needs specific conditions ( higher light exposition for example).
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Culture :
- Solid culture: we can use LED or Neon as a light source.
- Liquid culture: we need a set-up where we can control the illumination very well because we need much more light.
- Can the strain be cryopreserved ? Yes, with a mix of methanol and DMSO, cyanobacteria can be stored at -80°C.
- it’s better to use a microbiological safety cabinet for the manipulations
- Do we have the equipment suited to the specific cyanobacteria growth conditions? He advised us to look for available equipment at CRITT and TBI. He introduced us to key contacts in these two labs. Maybe we can find at TBI a bioreactor for eukaryotes that we could adapt to our cyanobacteria
Results
We had the chance to benefit from complementary training from our university in molecular biology for the UPS students, and in fermentation for the INSA students, as well as assistance from our instructors for any troubleshooting. However, we needed advice and counseling about unconventional lab material such as light panels for the cyanobacteria. The detection part of our molecules also necessitates people with experience in chromatography approaches. We were lucky to have this kind assistance from three technology centers in Toulouse (CRITT, ICEO and TWB).
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CRITT, technology center
We contacted Élise Blanchet, a member of CRITT, an innovation and technology transfer center. They have experience and material for cyanobacteria growth. She accepted to lend us some LED panels for cyanobacteria culture. She explained us that if the biomass increases too much, the lighting of our culture could become problematic, as light access can be very heterogeneous between cells. The CRITT help and experience was a great asset for us and reassured us with using cyanobacterium from scratch.
Subject : discussion of cyanobacteria materials
- CRITT has a broad knowledge of microalgae, they possess and master equipments that could be useful to us. At the moment they are not working on these organisms, so their materials are potentially available.
- They have test benches made of plastic bags that we could borrow. We may have to adjust them to our needs, they can be on site to help us with the installation. Neon lights can be provided to ensure the lighting of the cultures. However, the pockets of the test bench can hold up to 40 liters. This may not be suitable for our use, which at the beginning will be much smaller. The temperature cannot be regulated either, which is a problem for us.
- For the beginning of the project (where we would have a small volume of culture and petri dishes), they can lend us an oven with lighted agitation. We need to check if it is available.
- They can lend us 4 panels of modular LED Aquabeauty 70W intensity, we just have to check that they are in good shape to work with. We can place them in front of the ovens that we already have in the lab and also around our bioreactors.
- They also mentioned how culture would become more opaque as biomass increases. The cells in the middle of the culture would have less light or none at all, highlighting that agitation will be crucial.
TWB, technology center
Toulouse White Biotechnology is a demonstrator forging links between basic research and industry to help new biotech companies, it is also a platform with adequate equipment to perform services. At the end of our experimentation period, they helped us to analyze our culture supernatant samples in GC-MS. This allowed us to detect the molecules, and more particularly the ionones, that we had managed to produce with our engineered yeast strains.
ICEO, technology center
ICEO is a platform of engineering and screening for original enzymes directed by Sophie Bozonnet at Toulouse Biotechnology Institute. They helped us to analyze our molecules with a GC-MS. This allowed us to determine the best analytical protocols to detect the ionones we wanted to produce in our project
Application
Nowadays, more than ever, a good project has to be responsible, sustainable and safe for the planet, besides being economically viable and well perceived by the general public. This requires us to adopt the best practices, and to think further into the future of the project. As for the more “scientific” aspects, we had to integrate the ideas and knowledge of real experts and stakeholders from molecule extraction to commercialization, including ethical considerations and general public opinion.
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Carole Molina Jouve, Head of the Toulouse bioeconomy EUR
We talked with Carole about the extraction of our molecules. As we wanted to have a sustainable production, we excluded the use of solvents to extract our molecules. We had a look over extraction methods without solvents and we came across the pervaporation techniques. According to Carole, these methods could be very interesting for our project, as we could extract the molecule directly during the culture. She told us that there are different membranes to perform the extraction and that we could find specific membranes for our molecules.
Subject : downstream process
- For Carole, it is possible and interesting to use pervaporation in our project. This process is more promising than liquid-liquid extraction but also more expensive.
- Which In Situ Product Removal (ISPR) model do you recommend ? It is better to use the proposed 4th ISPR model because it allows to extract molecules during the culture. This is an advantage if the molecules produced are toxic for the cells.
- There are different types of membranes. Some work with pressure differences. The pressure gradient allows adsorption, transport of molecules and then desorption. Christelle Guigui is working on membranes and could brief us about the different existing membrane methods, and advise us on the most suitable for our project.
- How can we recover the odorous molecules we are interested in ? In pervaporation, the aromas may dissolve into ethanol during the culture, but she has heard of cases where they have not recovered the ethanol and it was not a problem. (knowing that ethanol is also used for the creation of perfumes).
- Can we use organic solvent ? Yes, however, it must have a very low solubility and must not interact with microorganisms. Is it “sustainable” ? Using organic solvents is not necessarily a problem as they can be recycled.
- Is it interesting to use methods with solid balls in order to extract our molecules ? Solid balls can be specific to our molecules. We have to be careful about compatibility with microorganisms. When we add balls in a medium, there can be damage to our cells due to friction
- It would be interesting to reconstitute a liquid medium with molecules of interest in the expected quantity, then to do a pilot test with a company in Toulouse, where we would put our molecules in a culture medium to try to extract them.This would allow us to test the extraction method.
Marie-Pierre Escudié, ethician at the Gaston Berger Institute
We contacted Marie-Pierre to talk about the ethical part of our project. The ethical part relate to both the fact that we use GMO, and the environmental impact of the perfume production. She told us that it would be great to compare the environmental and societal impact of the chemical versus the biotechnological method. She advised us to consider the limits of our project, to have a global vision of it and to be as realistic as possible.
General public, high school and UNADEV
Our project being based on synthetic biology, it was important for us to exchange with the general public to get their feelings on a perfume created from synthetic biology. The results encouraged us to continue, but also pushed us to take into account the standards defining the natural property of a product. We also noticed the lack of knowledge in both synthetic biology and knowledge of odors as well about the world of perfumery. We then embarked on a process of creating a board game we tested with the help of high schoolers and visually impaired people from the UNADEV association. These exchanges allowed us to better understand the needs of the public and the development potential of our project
Subject : Testing our game
- We test our game with 4 persons blind or visually impaired, between 30 and 70 years old, coming regularly to the association.
- When we arrived we knew that our cards were not adapted at all and we thought that the easiest way was to transcribe them into Braille. However, we were wrong. They explained to us that actually only 2 % of visually impaired people were able to read Braille. For young people it is easier to learn Braille but for people who become visually impaired later in their lives that’s more difficult, and most of them never learn. Also, for many reasons Braille is not adapted. For example people with manual work can have less sensibilities on the fingertips, so it is difficult for that type of people to read the Braille. Also, the texts on our cards were too long to be translated into Braille, they would become excessively large cards.
- For all these reasons they told us that vocal cards would be more suitable. They spoke to us about systems like “Pen frame”, which are actually pens with pre-record audio. The concept is to make a card with code that can be scanned by the pen and launch the audio corresponding to the card. In the same spirit, a system with a central box could read cards with codes and also serve as a vocal plateau.
- They gave us feedback about our perfume card. As we made small cylindrical counters to put on the cards to indicate if a smell was recovered or not, they told us that we could make the circles of the cards in relief so that we could position the counters correctly on the card. About these counters, for them the difference between the 2 faces, one smooth and one with an engraved lock, could be felt with the fingers. The color of the counters was not optimal. The counters are white, as the majority of the perfume cards, for blind people that don’t change anything, but for visually impaired people that could discern the 2, it can be difficult.
- The yeast counters can be improved too. We made our counters in 4 different colours but that was not optimal. It would be necessary to make 4 pieces of different shapes or to add different marks on the back of the pieces that can be felt easily with the fingers.
- We had our tray engraved on a wooden board. They could follow the engraving but it was not deep enough for them to recognize the boxes on the plateau. Actually that is easier for them to recognize a texture made in relief than an engraving. It would also be necessary to adapt the board with a system allowing clamping of the pieces into the squares.
- We brought dice with small engravings for the numbers, actually they could recognize the numbers on the dice but they would need larger dice to make it simpler.
- They also gave us an overall feedback on the concept of the game. For them it was interesting to mix knowledge of smells and biology. The level of the questions on the cards were perhaps not adapted to their level, they told us that they would think about making several levels of difficulty in the questions on the cards or maybe that the game could be played in teams.
- They were very excited about our game and our initiative to adapt it to visually impaired people. Actually there are not many suitable games at the moment. They were interested in helping us to continue adapting the game, retesting it and supporting us in the production.
- We also asked them questions about their sensitivity to odors. We thought that since one of their senses was impaired they might have a more developed sense of smell and that they might benefit from odor education. In reality not at all, they explained to us that they did not have a particularly developed sense of smell but rather that they were more sensitive to it. Not being able to see or see badly, they concentrate more on other things in the environment such as smells and are therefore more sensitive to them than seeing people.
Pierre-Alain Hoffmann, Kyanos
Pierre-Alain is a member of Kyanoss, a company working with phototrophic organisms. We contacted him to discuss the material we needed to work with the cyanobacteria. He told us to buy or to find a photometer to check exactly the lighting at the surface of our culture. We talked about other parameters of the culture like CO2 and temperature. He also helped us with the entrepreneurship part, especially to make our calculations for our production.
Subject : discussion of equipments dedicated to cyanobacteria growth
- At Kyanos, they work on microalgae with non-sterile and non-contained processes, they do not work with fermenters. Because of a recent geographical re-organisation, they are upgrading their fleet of equipment. They have 4 photobioreactors, 2 with a volume of 1.4 L and 2 with a volume of 5 L.
- Pierre-Alain pinpointed the need for warm white light, possibly enriched in red or blue. Our irradiance range corresponds to his advice.
- Quantifying the light with a photometer could be important to check the surface of the culture media. Irradiance is never homogeneous. He sent us references to calculate the amount of light necessary.
- A bioreactor with optional lighting, temperature control, etc. is very expensive. The solution is therefore a "fab lab" method which consists in using classical agitated and thermostatically controlled ovens for cultivation, with a window or a plastic cover to hang a row of LEDs. These leds, very cheap, must be controlled from the outside (it is necessary to choose a model with tunable intensity, and a mix between red and blue leds). He provided us some references.
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Regarding CO2 enrichment in the media:
- BG11 contains dissolved carbonate. It can be used as a source of CO2 for cyanobacteria. Be careful with BG11, it precipitates quickly, it is a complicated medium to prepare and use.
- Ask for help from the CRITT-GPE side: perhaps they could help us with the measurement of inorganic carbon.
- Growing 5% CO2 supplemented culture with a bubbler cylinder seems fine. Another possibility is to use medical mixers that are made to enrich air with oxygen but can work with CO2.
- CO2 is not the most difficult parameter to control, the temperature is
Robertet, international company
Robertet is an international company producing compounds for perfumes, cosmetic products and aromas for food products. We had the chance to present our project to Karine Kormann (raw material development manager), Anthony Pegard (perfumery development manager) and Thierry Roger (responsible for the sustainable development part of their production). We wanted to have additional feedback from industrials on our project, and also potentially find sponsors. Our project actually caught their attention and they see a real potential in its development. They supported us, became our main sponsor and brought us an invaluable assistance regarding the entrepreneurship part. They helped us with the standards used in the perfume industry and furnished upadted datas about the current market prices of raw materials, necessary for the dimensioning module of the entrepreneurship part. Eventually, a financial accord is in construction between Robertet and TBI for developing Elixio after iGEM. This discussion allowed us to realize how applicable our project could be for a company, and how important it was to communicate about this kind of biotechnology to democratize its use.
Subject : Perfume industry and industrial demand
- They enquired about our perspectives after the project (a company, a patent?) and about our instructors and resources. They were also curious to know how we heard about Robertet. We told them we found them on the internet as a company interested in biotech projects (especially with Gingko on the rose perfume).
- Thierry wanted to check with the legislation about using GMO and our approach with modified microorganisms.
- Another question was about our compounds toxicicity for the microorganisms.
- They wanted to know what kind of final product they would get if we initiate a collaboration (the culture medium or a "purified" extract).
- Thierry found the project very interesting. Extraction of natural compounds is very dependent on climate and plant diseases. Our solution allows natural production all along the year.
- Production of ionones in a natural way would be a definite advantage.
- About land occupation, Robertet grows violet for the green scents. Violet is a small plant growing at ground level. Its cultivation takes a lot of space and the yield per hectare is really low. With a bioreactor production, these surfaces would be free for other use. However, it could take years to set up this kind of production.
- Anthony proposed to produce the essence of violet leaf without using solvents.
- They proposed a partnership and would be glad to welcome the students for internships. They would like to participate as financial partners for us. They also agreed to let us film at Robertet for our video.
- They told us they were not so interested in the violet scent per se but by the proof of concept of proving the easiness to use synthetic biology to develop new perfumes
Conclusion
During the whole project, we met specialists who helped us on numerous and broad aspects. All these meetings allowed us to start from our basic idea, and to build a final realistic and applicable project. It made us understand how important it was to know how to network with experts in order to find solutions and get advice. This wiki page and others have been built with the very idea to transmit this precious information we received to future iGEMers.
We can only emphasize on the importance of meticulously defining every step of our project, and to find experts to transform each of these steps into a success key.
