Team:Toulouse INSA-UPS/Entrepreneurship

Overview



Our objective is to make our ELIXIO project industrial. In this entrepreneurship part, we will demonstrate that our project is feasible, scalable, and inventive. To do so, we first established the aims and missions of the future company as guidelines for our project to become industrial. Then, we imagined the milestones that we must reach as a company through a timeline. We also developed a marketing strategy to know which customers to target and which production means we needed. The establishment of these milestones and our marketing strategy have led us to conceive the building of a factory and to imagine a whole viable and sustainable industrial process. We made many predictions allowing the dimensioning of our industrial solution. Thanks to this dimensioning, we were able to calculate the production and sales costs of our industrial process. This led us to establish a business plan and its risk analysis.

Aims and missions of the company



The company's goal is to reproduce a violet accord using synthetic biology. The final objectives are to avoid the use of oil, arable land and to save energy (see the implementation part for more details on the production issues).

Building the aims and missions of a future company is a key step in the development of an industrial project. Thinking about these different issues allows to establish a line of conduct that the company will have to follow. This analysis also allows us to define the most important milestones to be reached for the product to become marketable. We have determined 5 critical steps in this process. They are outlined below, in decreasing order of priority.


Before the finished product (a violet accord) is obtained, the production chassis must first be designed (see design part) The yeast is used to produce the essential floral notes of the accord and the cyanobacteria the complementary green notes.


To make our product sustainable and with the lowest possible impact on the environment, we want to create a coculture between cyanobacteria and yeast. Atmospheric CO2 is implemented in the coculture to grow the cyanobacteria. Then, the cyanobacteria produces sucrose which is used by the yeast to produce our odorant molecules. Very few inputs are added to the culture for the production hence making it consistent with our bioeconomy-based project.


The market needs are important, and our production must be able to meet them. We must therefore adapt the various parameters of the coculture to obtain maximum production at minimum costs (see modeling and dimensioning parts).


The clients needs are specific and unique. To best meet the expectations of our potential customers, we would like to offer a versatile violet accord through a customizable system. The proportions of each odorant molecule in our accord can be fine-tuned. This requires determining which quantities of inducers allow obtaining which proportions of odorous molecules.


Natural and local products are increasingly popular with consumers. This is why we want our final product to be labelled as natural. Our manufacturing process will have to meet the specifications of the ISO 9235 (“ISO 9235,” 1997) and 16128 (“ISO 16128,” 2016) standards and eventually satisfies the demands of the COSMOS organic label (“COSMOS - Trust in organic and natural cosmetics,” n.d.).

ELIXIO in the future



The elaboration of the company's aims and missions gave us an idea of the initial requirements for our project. Here, we wanted to imagine the future of the company, once all these objectives are achieved. To give us a realistic idea of the temporal progress of such a project, we used the Photanol's timeline as a guide (“Photanol,” n.d.). Photanol is a biotechnology company that uses cyanobacteria to produce molecules of interest.

Thus, we believe that our violet accord could be commercialized in about ten years.

2022After the iGEM competition, the project continues with the same team as motivated as ever . ELIXIO becomes a start - up incubated in France at TWB (“Toulouse White Biotechnology,” n . d . ) . TWB is a research laboratory in biotechnologies linking researchers (pilot scale) and industry (pre - industrial scale) . 2021The iGEM Toulouse 2021 team give life to the ELIXIO project . The project aims to remediate the production of violet accord molecules through synthetic biology . During the development of the project within the framework of the iGEM competition, all the production stages are carefully analyzed . The CO 2 needed for the cyanobacteria should come from industrial effluents . The use of drinking water and energy must be minimized . 2025Pilot scale production in a 180 L flat panel is launched . The goal is to show that scale up is possible and that the project is industrially feasible . The initial team has grown, and experts have been recruited to go from pilot to industrial scale . In addition, the final strains were finely engineered and selected . 2026A strategic partnership with Robertet is established as they support our values and our vision of synthetic biology for a better world . The international company can provide us the financial and technical means that we might need . Thanks to this support, the pilot scale process has been further improved : production yields increase while energy consumption decreases . 2028A production cost of the violet accord was estimated thanks to the pilot scale . A high purity level for this accord was also achieved through the intervention of downstream process experts . An efficient and scalable pervaporation process has been developed to obtain a high purity product without any possible trace of GMOs . 2029The first industrial scale plant is launched and will demonstrate all our pilot scale estimations . The annual production amounts to 1 ton per year . Our product can also be considered as natural according to ISO 16128 and 9235 standards . In parallel, the former iGEM team is working on the production of other odorant molecules from the same chassis as to produce the violet accord in order to enlarge the company's fragrance palette . 2031The first batch of violet accord is commercialized! Our product meets the natural ISO standards and the various European safety criteria . The objective for the future is to open more factories around the world to recover CO 2 from industrial effluents . Moreover, the founding team has a lot of ambition and wants to innovate : the next product will be a hyacinth accord!

Marketing strategy



Market segmentation for natural fragrance ingredients


We have segmented the synthetic biology fragrance market to better assess our competitors and define our target.

The diagram below (Figure 1) shows the different companies in the market. This list is not exhaustive, but it helps to better understand the structure of this market.

Figure 1: segmentation mapping

The leaders in the fragrance ingredients market are versatile and often produce so-called "synthetic" ingredients as well as natural ones, including biotechnological approaches. These ingredient specialists can work in collaboration with other biotech companies (such as Robertet and Gingko). Indeed, these companies are specialized in biotechnology in the broadest sense of the term (fermentation, etc) which offer ingredients for several sectors of activity (cosmetics, health, food, etc.). They are not necessarily specialized in the production of ingredients for the perfume industry and the use of synthetic biology in particular still seems marginal.

Target definition


Biotechnology companies have already offered their help in providing natural ingredients through bioprocesses, but they are not necessarily specialized in the manufacture of fragrance ingredients. Perfume and cosmetic companies buy their ingredients from companies such as Givaudan, Firmenich, Symrise, IFF or Robertet. It would therefore be more relevant to touch these ingredient suppliers directly. Leaders in the ingredients and fragrances market have started to turn to biotechnological approaches to manufacture certain ingredients. This is why we decided to focus on medium-sized companies that may not yet have been able to explore synthetic biology to meet their needs.

At the beginning of our activity, we will not be able to produce sufficiently to meet the general market needs for ionones for example. This is why we decided to have only one target to start with: a world leader in natural ingredients, Robertet, which has expressed a need for additional naturalness for certain ingredients, and which has been following the ELIXIO project since the beginning.

We have chosen to do Concentrated Marketing because we want to position ourselves as a specialist. We focus on one market segment and therefore only offer one marketing plan.


Positioning strategy


Figure 2: Order of magnitude of purchase prices according Robertet

Our final product will not be able to economically compete with its chemical counterpart. But in view of the challenges of sustainable development and the strong demand for naturalness in recent years, some customer segments are willing to pay more for a better quality and more sustainable product. Thus, our positioning strategy is vertical differentiation through perceived quality.



We differentiate ourselves from other companies through:

Our added value:

  • Ingredient produced by synthetic biology

  • Mute flower ingredient production

  • A system scalable/modulable to the customer’s needs

  • A very low carbon impact, possibly the lowest

  • No mixing of isomers, as found in chemical processes

  • Products available all seasons

Dimensioning



Introduction


The objective of this document is to estimate the energetic, economic, and ecological costs if our project would become industrial. With the help of our sponsor Robertet, we estimated that an annual production of 250 kg of violet accord could meet their needs. By using our modeling part, we could estimate that an industrial installation based on our process could produce 260.5 kg of violet accord per year. This way, we can meet the needs of our only customer, Robertet, with even a small margin in case of production issues during the year. This preliminary study provides an overview of the financial and ecological impacts of our project, thus driving our entrepreneurship vision when maturating the project.

We would like to specify that this study is meant to be as exhaustive as possible. Thus, we have answered the maximum of problems that could arise from the launch of a start-up. However, we are aware that some aspects, such as the purchase of quality control equipment that would require a more detailed study to determine the tests to be performed, are not covered by our study. The dimensioning allows us to have a coherent order of importance of the various costs associated with the industrial process.

In a first step, we chose our industrial installation, i.e., we dimensioned the reactors necessary for the desired production. Then, we calculated the energy costs related to this production. The calculation of the energy consumed by the bioprocess allowed us to estimate a price and the CO2 equivalent produced by our reactors. Once the molecules of the violet accord are produced, they must be extracted from the culture medium. So, we have studied the costs of pervaporation and the other steps related to our production process such as waste treatments. Finally, we have studied the economic aspect of our industrial project. Expenses and investments are important in any industrial set up and were also estimated. All this analysis eventually allowed us to calculate the production cost of our violet accord as well as the environmental impact linked to the industrial process.

To visualize each step of the production process, we have established a flowsheet of our industrial installation (Figure 3). We will discuss in the following the dimensioning of the choice of the reactors and the different parameters allowing optimal growth conditions for the two microorganisms.

Figure 3: flowsheet of the industrial installation

Later in this section, the electrical costs and the equivalent CO2 emission will be estimated from the electrical consumption of each element of the installation. In France, the cost of the kWh for an industrial is €0.1469 including tax (“Fiche descriptive - Offre de fourniture d’électricité au ‘Tarif Bleu,’” n.d.). This index gives an idea of the environmental impact of energy consumption. In France, the CO2 equivalent of 1 kWh of electricity is 33g (“Bilans des émissions de GES - EDF - France métropolitaine,” n.d.).

Upstream and fermentation process


Design of the PBR to achieve the desired production

There are many different types of industrial photobioreactors, with their pros and cons. The thickness of the reactor walls should be as small as possible to ensure the best possible light penetration. Since the reactor is thin, solar energy is sufficient to maintain an optimal culture temperature. However, in summer or winter, the outside temperature can be far from the desired condition. To overcome this problem, the reactor can be immersed in a pool of temperature-controlled water (Huang et al., 2017; Morweiser et al., 2010). The choice of tank material is also crucial. Glass sides are ideal for small photobioreactors while plastic sides (PVC, PET...) are more suitable for larger reactors (Huang et al., 2017; Johnson et al., 2018). This choice must consider a great lifespan of the reactor and an easy maintenance.

Type of existing photobioreactor (PBR)

Figure 4: column airlift PBR (Mohler et al., 2019)

  • Column airlift: a column airlift can be in the shape of a solid or hollow cylinder to allow a better light homogeneity. Aeration is provided from the bottom of the column. This type of PBR does not consume much energy and the mass transfer is optimal. The maximum dimensions of such a reactor are however limited due to structural constraints. They are also not easy to clean and require a significant initial investment (Huang et al., 2017).

  • Tubular PBR: here, the culture takes place in circular pipes with a diameter of less than 60 mm. A pumping system is used to aerate and mix the culture. A very high amount of energy is used by the pumping system, up to 200 W/m3. This type of reactor is very effective but also expensive in terms of auxiliary energy (Morweiser et al., 2010). In addition, the culture medium is not very homogeneous and maintenance is complicated in such PBRs (Huang et al., 2017).

Figure 5: tubular PBR (Alaswad et al., 2015)

Figure 6: plastic bag PBR (“Food-themed art installations to be served up at LA parks,” 2019)

  • Plastic bag: big plastic bags, with water as support or fixed to a metal stand, can also be used for the culture of cyanobacteria. The materials of the reactor are cheap which make this system to be very economically attractive for mass production (Morweiser et al., 2010). However, this type of PBR has many disadvantages. Due to their thickness, they can easily break, agitation is not properly ensured, and the lifespan of a plastic bag is very short (Huang et al., 2017).

  • Flat panel airlift: this reactor can be used in a long period of time because it is made of glass and concrete. The cleaning and the maintenance are simple which also contribute to its long lifespan. Moreover, this reactor shows good performances: it has a high ratio surface-volume which provides a significant amount of light, the shear stress on the cells is minimal while a good mixing is ensured thanks to its geometry, the energy consumption is minimal compared to the others PBR design… All these advantages make the flat panel reactor the most promising for an industrial production (Huang et al., 2017).

Figure 7: flat panel airlift PBR (“Product Cultivation Module – SUBITEC,” n.d.)

Choice and dimension of the industrial PBRs

Because the flat panel is the most cost-effective PBR, we decided to choose a flat panel photobioreactor for our industrial co-culture (Huang et al., 2021, 2017; Lindblad et al., 2019).

To produce 250 kg per year of violet accord, we opted for an installation of 40 PBRs of 180 L proposed by Subitec (“Product Cultivation Module – SUBITEC,” n.d.). After negotiations with this manufacturer, the price for the 40 PBRs should be about €250,000. This dimensioning of the photobioreactors will be used as a basis for the calculations of the modeling part. Moreover, the results obtained with the model will allow to estimate the various economic and energy costs of the process.

PBRs = €250,000
Lifetime of the PBRs: 20 years. So, with an amortization on 20 years: €12,500 per year for the PBRs


Sterilization of the reactors

The reactor is sterilized and cleaned with a 35% hydrogen peroxide solution. According to the publication of Derwenskus et al., 2020, we would need 7.41 cubic meters of hydrogen peroxide and 215 cubic meters of water per year to sterilize our 40 reactors. The associated expense would be €5,116 per year.

PBRs sterilization = €5,116 per year


Power requirement for the fermentation process

Heat exchanger

To save energy, it would be interesting to use solar energy to heat and light the photobioreactors. For this reason, the reactors should be placed outdoors. Moreover, the temperature regulation can be done with heat exchanger. By installing our plant in France, on the Mediterranean coast, we can maximize solar lighting and use seawater as a coolant fluid to reduce the use of clean water in our process. In addition, Robertet's production facilities are also located in the south of France. Thus, we could minimize transportation by locating our factory in the same region. In addition, it would be possible to use the CO2 emissions from Robertet's plants to feed our installation. These industrial effluents would thus be recycled.

In order to use as little non-renewable energy as possible, we want to heat the water of the heat exchanger with thermal panels, thus with the available solar energy. Moreover, to reduce the consumption of drinking water, we want to use sea water to regulate the temperature of our culture. We estimate that the temperature of the sea water is 20°C on average and we want to heat this water to 40°C. The water will be stored in a 1,000 L tank.

Quantity of energy required to heat 1,000 L of water:

\( Q=m_{water}cp{water}(T_F-T_I)=1,000*4,180*(40-20)
\leftrightarrow Q=83,600,000 J=23,200 Wh \)

This tank will be heated once a day, so 365 times a year:

\(Q_{year}=8,476kWh/year\)

In the south of France, it is estimated that an average thermal panel can produce 500 kWh.m-2 per year and the cost of a square meter of thermal panel is about €1,700 (“Panneaux solaires thermiques : Guide d’achat - Conseils Thermiques,” n.d.). Thus, it is possible to determine the surface of thermal panel necessary for our installation and the associated cost.

It must be considered that solar panels do not have a 100% efficiency. Thus, taking a yield of 80% (“Le rendement des panneaux solaires thermiques et photovoltaïques,” n.d.), the necessary surface is:

\(\textbf{Thermal panel surface}=\frac{8,476}{500}*1.2=20.3 m^2\)
\(\textbf{Associated price}=20.3*1,700=€34,510\)

The sea water is stored in two separate tanks and the associated price for these tanks is €989.

Solar panel (20 m2) = €34,510
Lifetime of the solar panels: 20 years. So, with an amortization on 20 years: €1,700 per year for the solar panel


Sea water tanks = €989
Lifetime of the tanks: 20 years. So, with an amortization on 20 years €49 per year for the sea water tanks


Lightning

On the French Mediterranean coast, the average monthly sunshine is 267 hours, or 3,204 hours per year (“Ensoleillement et climat sur Nice - Mairie de Nice et sa ville,” n.d.). It is necessary to light the PBR the rest of the time.

\( \textbf{Time of lightning per year}=365*24-3,204=5,556hours/year \)

The lamp we have selected consumes 560 W (“GC Bar 8 - Greenception,” n.d.). To light the whole surface of a flat panel, we need two lamps, so in total we need 80 lamps to light the 40 reactors.

\(\textbf{Electricity consumption of the lamps}=0.560*80*5,556=248,909kWh/year\)

One lamp cost €1,849 so for 80 lamps we must invest €147,920.

80 lamps = €147,920
Lifetime of the lamps: 5 years. So, with an amortization on 20 years: €29,600 per year for the lamps
Electricity consumption = 249,000 kWh/year


Agitation and aeration

The agitation inside the flat panel is ensured by the aeration. Air injection is provided by air stones. The size of the bubbles is reduced which increases the transfer of CO2 to the liquid phase and the agitation (Falinski et al., 2018). You can see more details about this choice in the modeling section. The flow of air enriched with CO2 must be sufficient to move the entire mass of culture medium. We chose an air compressor to supply our reactors. One compressor can supply three reactors, so we need 15 compressors for our entire installation. The compressor has a power of 1.07 kW (“KK70 - Compresseur sans huile by DÜRR TECHNIK | DirectIndustry,” n.d.) and they are used the whole year, i.e. 8,760 h.

\(\textbf{Electricity consumption of the compressors}=1.07*15*8,760=149,598kWh/year\)

Electricity consumption = 149,600 kWh/year


Pumps

Finally, the installation also requires pumps to fill the reactor with culture medium, to pump the cooling water and the medium loaded with odorant molecules to the filters of the pervaporation. The specifications and number of the different pumps are listed in the following table.

Figure 8: pumps electrical consumption and equipment cost

The heat exchanger, and thus the pump for filling the seawater tank, is only needed half of the year, i.e., 4,380 hours per year. To regulate the temperature of each reactor as well as possible, one pump is needed for one reactor, so we need 40 pumps in total. The pump for filling and emptying the reactor is used at the end of each batch, i.e., 30 times a year, because in one year 15 batches can be performed. Estimating that the filling and emptying of the reactor lasts 8 h in total, the pump is active 120 h per year.

Equipment cost = €14,515
Lifetime of the various pumps: 20 years. So, with an amortization on 20 years: €725 per year for the pumps.


Electricity consumption = 125,330 kWh/year


Culture medium

The culture medium is coyBG11 which composition is detailed in the publication of Hays et al., 2016. To perform 15 batches per year, we need:

\(\textbf{Volume of culture medium}=15*40*180=108,000L\)

Based on the publication of Derwenskus et al., we can estimate that the price for such a quantity of culture medium is €15,808.

The BG11 cannot be sterilized with a temperature increase. In order to eliminate 99.9% of all possible microorganisms (viruses, bacteria, fungi...), a UV lamp irradiating 330 Ws.cm-2 must be used (“La stérilisation UV de l’eau,” n.d.). The surface of one of our PBR is 3.6 m² and 1 m2 is equal to 10-4 cm2.

\(\textbf{Energy for 3.6 m²}=\frac{330*3.6}{10^{-4}}=11,880,000Ws\)
\(\leftrightarrow \textbf{Energy for one PBR}=\frac{11,880,000}{3,600}=3,300Wh\)
\(\leftrightarrow \textbf{Energy for 40 PBRs}=40*3,300=132,000Wh\)

As we do 15 batches per year, we need to sterilize the medium 15 times per year.

\(\textbf{UV sterilization energy per year}=15*132,000=1,980,000Wh/year=1,980kWh/year \)

In addition to UV sterilization, the culture medium is filtered. The filters are changed between each batch and a filter costs €422.39 (Derwenskus et al., 2020).

\(\textbf{Filter cost per year}=15*422=6,330€/year\)

Culture medium = €15,808 per year
Filter = €6,330 per year
Energy consumption = 1,980 kWh/year


Downstream process


Once the odorant molecules are produced in the reactor, they must be extracted and purified. This extraction will be done continuously throughout the culture using a pervaporation system. Since we will perform batch operations, we must treat GMO waste and wastewater at the end of each operation. These treatment operations have an economic and ecological cost that must be considered.

Extraction and purification of molecules

To continuously extract and purify the fragrant molecules, we have chosen a process of pervaporation. Indeed, the pervaporation allows a great energy saving compared to the distillation process classically used in the industry (Castel et al., 2020). Based on the publication of Castel et al., we can estimate that the extraction of 250 kg of violet accord consumes 120 kWh.


Energy consumption = 120 kWh/year


Waste recycling

Biomass waste (GMO)

According to the High Council for Biotechnology (Haut Conseil des Biotechnologies), genetically modified organisms only need to be inactivated to be considered as common waste (“Manuel du HCB pour l’utilisation confinée d’organismes génétiquement modifiés,” n.d., p. 24). Once inactivated, this waste can be used as fertilizer for example.

It is possible to estimate the mass of biomass to be treated per year thanks to the simulations of the modeling part. The concentration of yeast in the coculture is 2.5 g.L-1 and 40 g.L-1 for the cyanobacteria.

Figure 9: biomass production during violet elixir production

GMO waste is processed by an external company at a cost of €2,000 per ton.

\(\textbf{GMO waste treatment cost per year}=2,000*4.59=€9,180\)

GMO waste treatment = €9,180 per year


Wastewater

The amount of wastewater generated by the annual production is equivalent to the volume of culture medium used, i.e., 108,000 L. Once the biomass is removed, the culture medium can be used in addition to the seawater in the heat exchanger. It does not contain any hazardous compounds and therefore can be safely discharged into the sea.

Wastewater = 108,000 L


Investments


To launch our production of violet accord, we need to invest in equipment, manpower and the building of a factory. All these investments are amortized depending on their nature and their cost must be considered in the final production cost of the fragrance. This part will allow to estimate the production cost of 250 kg of violet accord as well as its selling price. In addition to the economic costs related to the project, we wanted to evaluate the environmental impact of an industrial production.

Calculation of expenditure and investment

Labor costs

The minimum wage in France is €11.6 per hour for a manager (“Le coût d’un salaire au SMIC avec les charges,” 2021). To ensure uninterrupted production, 11 employees working 35 hours a week are required. Removing public holidays and weekends, it is estimated that employees will work 250 days per year, or 6,000 hours.

\(\textbf{Labor cost}=11*6,000*11.6=765,600€/year\)

The batches should last 3 weeks, and 3 days of maintenance are necessary so 24 days in total. Counting 365 days per year, it will be possible to realize 15 batches per year.

Labor cost = €765,600 per year


Land and factory construction costs

We want to set up our factory by the sea and in the PACA region (France) to be as close as possible to Robertet's factories. The average price for 3,000 m2 of industrial land is €1,012,500. The construction of a permanent factory requires material and manpower which represent €1,000 per square meter. Finally, for a factory of 2,000 m2, the cost is around €2,000,000 (“Quel est le prix au m2 de la construction d’un bâtiment commercial ?,” 2019).

\(\textbf{Land and factory construction cost}=1,012,500+2,000,000=€3,012,500\)

Land and factory costs = €3,012,500
Lifetime of the construction: 30 years. So, with an amortization on 30 years: €100,000 per year for the land and factory


Pervaporation equipment costs

Based on the publication of Derwenskus et al., we can estimate some equipment costs. The cost of installing the pervaporation process can be estimated at 15% of the total infrastructure cost (Castel et al., 2020), i.e.:

\(\textbf{Pervaporation investment}=0.15*3,012,500=€451,875\)

Pervaporation installation cost = €452,000
Lifetime of the installation: 30 years. So, with an amortization on 30 years: €15,000 per year for the pervaporation installation


Biorepository cost

One of the most important risks for our company is to lose the strains producing the violet accord. To avoid this prospect, which would put an end to the business, the modified strains must be stored properly at -80°C. The price of two such freezers is €17,498 (“Congélateurs-coffres à ultra-basse température de -86°C, RevcoTM série CxF,” n.d.).

Freezers cost = €20,000
Lifetime of the freezers: 20 years. So, with an amortization on 30 years: €1,000 per year for the 2 freezers


Economic and ecologic evaluation


Water consumption

In France and more precisely in the PACA region, 1 cubic meter of water costs €3.70 (“Le prix de l’eau,” n.d.). In our process, we need 108,000 L of drinking water per year, which is a cost of:

\(\textbf{Water cost}=108*3.70=€399.6\)

Water cost = €399.6 per year


Environmental cost

The electricity consumption allows us to calculate the economic and ecological costs (via the equivalent CO2) related to our process. As a reminder (introduction in France, 1 kWh costs €0.1469 and generates 33 g of CO2 equivalent.

Figure 10: electrical consumption of the industrial installation

The energy required to operate our industrial process would generate 17 tons of CO2 equivalent per year. However, the growth of biomass throughout the year has a strong impact in terms of CO2 absorption. Thanks to the results given by the model, we can estimate that the biomass should convert 8 tons of atmospheric CO2 into the product of interest. This means that almost 50% of the CO2 emissions produced by our process are absorbed by the biomass. To sum up, our process rejects only 9 tons of CO2 equivalent per year, i.e. 34 kg of CO2 equivalent per kg of violet elixir. By way of comparison, a quarter of the way from Toulouse to Paris by plane emits 34 kg of CO2 (“Mon Convertisseur CO2,” n.d.). In the future, the energy consumption should be reduced in order to aim at a zero-carbon emission process.

Waste management, culture medium manufacturing and the chemicals used to sterilize the reactors also have an environmental impact. However, this impact is more difficult to measure than the energy cost. Our objective is to minimize this environmental impact, and we want to make the most of industrial effluents. First, since our cyanobacteria consume CO2, we can capture emissions from a factory to valorize this carbon. Then, after sterilizing the GMOs, the dead biomass can be used as fertilizer or compost.

In order to have a coherent point of comparison between the different production methods (natural, chemical or biotechnological), it would be interesting to carry out the same study on the two other production processes.


Electricity cost = €77,000 per year

Production cost

In the table below, the different costs mentioned previously are summarized. In the end, according to the depreciation and electrical costs, it is possible to estimate the annual economic cost to produce 260.5 kg of violet accord.

Figure 11: detail of the production cost

Conclusion


Figure 12: distribution of the yearly cost of production. As a percentage of production cost: 73% labor cost, 10% land and factory construction, 7% electricity, 5% other, 3% lamp, 2% photobioreactors.

A production cost to be optimized

The annual production cost is €1,047,000. Therefore, the price associated with 1 kg of violet elixir is €4,019. This selling price is much higher than the data provided by Robertet (€350 per kg of non-traceable "natural" product and €10 per kg of chemical product). For our process to be viable, research and improvements must be made to reduce this selling price. The most obvious improvement is to increase the production capacity of our strain. There are two ways to do this, as indicated in the modeling section:

  • by decreasing the yield of sucrose production it is possible to increase the production of odorant molecules. The cyanobacteria strain should be engineered in this way.

  • by promoting the transfer of CO2. It is by modifying the shape of the photobioreactors that the transfer of CO2 could be optimized and the production of odorous molecules as well.

In fact, increasing by ten our production capacity would allow us to compete with the so-called natural product. Another possibility is to reduce our expenses by automating some tasks of the production thus reducing the costs linked to the manpower.


A valuable production despite its cost

However, our product has many advantages over other means of production and these differences could still allow us to sell our product at a higher price. Indeed, our production is ecological, natural, and transparent. Robertet's customers generally have very strict specifications, particularly concerning the origin and production methods of the raw materials. Even though our manufacturing process is more expensive than chemically synthesized and natural molecules, the transparency and natural appearance of our product is a great asset for Robertet.


Beyond the violet, a proof of viability for other molecules

Finally, our project is a proof of concept, and here we show that we are not far to be economically viable. There are many scent molecules which could be of a great interest to produce using synthetic biology and biotechnological processes. Some of these molecules are rare and cost-effective enough to allow to ensure the interest of the synthetic biology approach.

Business plan



Business Model Canvas


We made a Business Model Canvas that allows us to present our business model clearly. It allows us to define our priorities, the steps to be taken and the ways of improvement.

Figure 13: Business model canvas

Our project has a real added value and meets an actual industrial need. We are fortunate to have a close relationship with our client even before the company was created. The search for new customers is therefore not a priority for the moment. Moreover, the iGEM network and the knowledge of the researchers in the TBI laboratory will be a real intellectual support to progress in our research. However, the creation of such a structure requires significant fundraising. Despite the support of the industrialist, it will be important to consider setting up decent financing to complete the revenue streams.


To begin with, it is imperative to consider the choice of legal status for the company, but also how to protect our scientific work. We also need to find out about the legislation to label our product as natural.

Choice of legal status


In France, the ideal legal status for a start-up is the SAS (Société par Actions Simplifiées). It is flexible and easy to create. A minimum of 2 partners is required, but no minimum amount of share capital is required. The partners can either make a cash contribution or a contribution in kind (“Start-up,” 2016). The founders will also be able to determine how decisions are taken at the meeting.

Creating an SAS generates costs but these are negligible in view of the overall investment. There are compulsory administrative costs which include the expenses of legal notices and the remuneration of the clerk for the registration service (€295). Other expenses are to be expected such as the fees of a contribution auditor, trademark registration fees, for the blocking of share capital, and costs for support for creation (up to €2,500).

Figure 14: expenses associated with the creation of an SAS

It is necessary to plan between €300 and €5,000 of costs to create a SAS.

Intellectual property


In France, the INPI (National Institute of Intellectual Property) is the body that manages all applications for intellectual property titles. Improvements of our already implemented method for fragrance production in yeast would be of capital importance for patent submission. Moreover, our goal is to be able to extend our methods to new odours. Patent protections would then be possible. The filing of a patent allows a monopoly of exploitation for a maximum period of 20 years. It confers a right to prohibit any use, manufacture, import, etc., of our invention made without our authorization (“Une protection efficace de vos innovations techniques,” 2015). It is a very important element in gaining credibility with our partners and clients. The filing of a patent costs around 700 € (“Combien coûte un brevet ?,” 2015).

It would be interesting to register a trademark too. It is the only intellectual property that can never fall into the public domain since it is indefinitely renewable (to be renewed every 10 years). We have verified the availability of our trademark in the INPI database: our Elixio mark does not reproduce or imitate a sign that enjoys a prior right for products or services, or activities that are identical or similar to ours. Electronic filing of a trademark on INPI costs €190 for a product or service belonging to one class. You will have to add €40 for each additional class (“Combien coûte une marque ?,” 2020; “La propriété intellectuelle,” 2017).

ISO Standards


Our objective, to distinguish ourselves from our competitors, is to be able to label our product as natural. For this, the criteria of two ISO standards must be met:

  • ISO 16128 (“ISO 16128,” 2016): according to this standard, "natural ingredients are cosmetic ingredients obtained [...] from micro-organisms". Moreover, the molecules produced must exist in a natural state. Our violet accord meets all these criteria.
  • ISO 9235 (“ISO 9235,” 1997): this standard defines as natural raw material a material "of vegetal, animal or microbiological origin, as such, obtained by physical, enzymatic or microbiological processes or by traditional preparation processes". By following this definition, it is possible to qualify our product as natural.

The fact that we meet the criteria of two ISO standards shows our prospective customers a guarantee of quality and transparency. Moreover, unlike chemically produced molecules, our production process is natural and ecological.

Risk analysis



Here, we have studied the risks associated with our project using the FMECA (Failure Modes, Effects and Criticality Analysis) method. This method makes it possible to anticipate the various risks associated with the project and to prevent them. It allows to prioritize the risks to know in advance which ones are to be treated quickly or are eventually negligible.

List of risks


Figure 15: list of risks and details

Frequency and criticality of these risks


In order to classify our risks, we have established a reference table for risk criticality calculations.

Figure 16: reference table for FMECA risk calculations

Figure 17: risk assessment with the FMECA method

As we expected, environmental risks are the most critical. Waste, especially GMO waste, can be complicated to handle while meeting the required safety standards. In addition, none of the iGEM students have experience in entrepreneurship. This lack of experience must be taken into account but should not be a barrier to our project: we are able to learn quickly.

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