The implementation of the project was divided into distinct steps following the development of an operations workflow to guide each one of them. In the first step, the team partnered with fruit producers to acquire juice production residues as those wastes are usually discarded. In the second step, with the residues (recycled into culture media) and the genetically engineered organism, we will produce bacterial cellulose in a more efficient way. We will be able to reduce the required time for achieving the stationary phase and also increase the conversion ratio of glucose to cellulose. They could produce specific products on demand for the final Consumers. One of our company’s main customers, hospitals, is a great example of how the implementation would be held as we would provide a kit with gels or cellulose sheets to be used in burn injuries (Cellulopolis BC kit). Research and development (R&D) customers can also order specific cellulose shapes to use as cell scaffolds. Furthermore, we can actuate as a supplier to companies who want to use cellulose as a base for other products such as cosmetics and pharmaceuticals.
The Cellulopolis project aims to act in two ways, on one hand, it delivers the idea of providing the kit quickly and effectively to industries in general, and on the other hand, it provides the product ready to its final consumer. We do not intend to deliver our genetically modified organism (GMO) at any time as we put in the effort to provide biosecurity efficiently, but rather a product of this GMO, which pharmaceutical and cosmetics producers can use in their processes, improving the manufacturing of their goods, this way the end consumers can make direct use of the products originated from our project.
For the distribution of products originating from Cellulopolis, a flow of operations was developed, aiming at maximum optimization and use of available resources. Brazil is a major producer of agriculture products and at the end of this production, there is a large accumulation of waste, which has no destination and ends up being disposed of incorrectly, harming the environment, and it is precisely at this point that the flow of Cellulopolis has its beginning. In cooperation with the fruit growers, we collect these residues, and using this material, we generate a culture medium for the cultivation of our bacterium, Komagataeibacter, which has shown to grow as efficiently in this medium as in others, commonly handled in the laboratory. In a laboratory environment, these culture media are made using these spare parts, for the growth of genetically modified strains, built by Unicamp_Brazil, after the growth of bacterial cellulosis, the purification of this is carried out, which will finally be made available in an expression kit, and scaffold production. This product will facilitate the treatment of burns with greater efficiency, in addition to making production cheaper, and meeting the most diverse demands, in addition to burns, can also be implemented as a fibrous scaffold for the skin, due to its properties. The flow of operations ends precisely at the delivery to final consumers, which can be hospitals, ordinary people, or the most diverse industries, such as pharmaceuticals and cosmetics, and can help to make new research in these sectors viable.
In the midst of the implementation, it is important to emphasize that this aims to be carried out in an economic and sustainable way, always seeking to deal with affordable prices, since in addition to being a more effective product, it also aims to reduce costs for the final consumer.
The team knew that to avoid any contaminants that could compromise the final consumer and to make the implementation of our product possible, it was extremely important not to make the microorganism available directly to citizens, but to its final product (The Cellulopolis kit). We took care to analyze the growth of biomass and cellulose production to make sure that the reproduction of microorganisms would be regulated by the production of the material assigned to them.
But the biggest concern of the team in terms of safety was that, since we wanted to make our product as accessible and with as few barriers as possible we took care to use only white-listed organisms in all of the steps of our process, that means that from the beginning of the product until it’s the end we would be working with organisms that were safe for human consumption.
As a scale-up implementation for the chassis organisms, the plan is to develop and implement a double kill switch System to control the reproduction of the bacteria and its spread. The first kill switch would be a nutrition-modulated one and the second kill switch would be a toxin-antitoxin system-based one. This way the Safety of the bioreactor, plant, and overall process of production would be
As challenges, we can mention the great handling difficulty that the Komagataeibacter bacterium provides. This organism takes 3 days to grow, which delays the process a little depending on demand and urgency. Komagataeibacter is a reasonably expensive bacterium to maintain, because its culture medium, HS (Hestrin; Schramm, 1954), commonly used in laboratories, carries a large amount of glucose, being 50g, while most common media takes around 2g - 4g. It is also valid to bring the complication in automating the removal of the cellulose blanket, after its production, in addition to the wide obstacle encountered in making this whole process, of cellulose production, become continuous, for reasons of possible contamination and also the impasse encountered at the time of removal of the cellulose produced, considering that when this polymer is removed, the cell is also removed.
We focused primarily on reducing times within the production system as well as response times from suppliers and customers, thus our process of choice was utilizing the 5 principles of manufacturing (commonly referred to as lean manufacturing). This process was very useful to make us understand how we would conduct our manufacturing processes and to always keep aiming for perfection. most of these steps were supported further by data collected in our process and market research. The challenges we will face in each of these steps will be mapped and organized in a way that mistakes can be fixed as quickly and as efficiently as possible.
To evaluate the technical and market feasibility of our product, we developed a simple work plan involving contact with biopolymer experts and companies with market experience in commercializing these products, in order to verify whether the kits thought for expression and scaffolding of bacterial cellulose are truly viable, and if not, how we could remodel our work to meet the commercialization requirements. For a more detailed description, visit the Entrepreneurship page.
According to the prospect made in the market analysis and in the strategic planning of our business (see entrepreneurship), our clients are corporations involved in the pharmaceutical and cosmetics industry mainly. However, our customers are different from our end consumers for a number of reasons. Since our customers are companies that would use our product in their services, our true end consumers are the people who use these companies' services. For example, if our bacterial cellulose is sold to a hospital that uses it to treat burns of a patient, our customer (who bought our product) would be the hospital, but who would really be consuming this would be the patient (hospital customer).
In this way, we define our end consumers as the clients of our clients. People involved in accidents with burns in low-income hospitals, who would make the best use of our products sold at low cost, or consumers of cosmetics containing our cellulose (purchased by one of our customers). With our cellulose production we can deliver the final product to hospitals, companies, and laboratories in the form of gel, ointment, scaffold, blocks or other conditions required by the customer.
The manufacturing implementation process was crucial from the beginning of the project as the team planned to make this project as feasible as possible. The initial bottlenecks of the cellulose manufacturing process posed some challenges to the overall production. Especially in the yield of each batch. Therefore for the next implementation steps, the team will apply the safety measurements and organize every key part of the manufacturing process, taking into account the risks and possible wastes we will have to manage on our way to perfect our product.