Our probiotics can produce enough vitamins!
In order to evaluate how much vitamins our probiotics could produce in idli during an overnight fermentation, we made a predictive mathematical model, taking into account the growth rate of micro-organisms in the idli batter and the rate of vitamin production, as well as the lag and the stationary phase of microbial growth.
Our diagram shows the amount of vitamin A, B2 and B12 that our micro-organisms would produce in a 12 hours fermentation. According to our model, adding 10 mL of saturated culture in the idli batter before the fermentation would be enough to meet the daily intake recommended by the FAO for those three vitamins.
VIDEO - Probiotics that produce vitamins in your food
We made a short animated video to explain the science behind our project - what are fermentation and synthetic biology, and how we intend to use both to fight malnutrition in an elegant and practical way.
Our iGEM experience
Before coming to the TFF Challenge, we presented our project at Boston last September, for iGEM - the international competition of synthetic biology!
It was an awesome experience. We learned so, so much, and we got to meet teams and specialists from all over the world! There were more than 260 teams, with as many incredible projects!
We presented our project in front of judges and other participants, and were proud to receive a Gold Medal, and to be nominated for the Best Food and Nutrition Project, the Best Integrated Human Practices, and the Best Presentation.
It was also a great opportunity to get feedback from scientists and other people interested in our project!
If you want to know more about it, you can go to our wiki, the website where we described all our thinking process, our experiments and our results over the summer: http://2015.igem.org/Team:Paris_Bettencourt
Manufacturing: the VitaCube
Our project is designed for poor regions: the cheaper our product, the more people it can reach. We didn't want to build labs in India, buy media and grow our strains in expensive structures. Therefore, we had to think of a cheap solution to grow the strains in a homemade media, made of common ingredients, cheap and readily available.
We made a series of experiments to see which strain could be grown in home-made media, and we discovered that all our probiotics were growing simply in water that had been used to cook potatoes or rice. Nothing else was needed, there is enough nutrients in just this cooking water.
It means that once someone has the probiotics, they can grow them at home easily and with no cost at all.
We also thought about how to distribute those strains to the population in the first place. We could have just lyophilized the strains but our goal is to design something cheap and easy to do for the locals, using only ingredients they have access to and not time consuming.
We realised a powder wasn't the best way to distribute our strains. We found better to make portions, easy to pack, with the possibility to pack several portions together. Portions must be easy to stock.
The most efficient and ergonomic shape appeared to be a cube. Moreover, the cube will mainly be added to Idli, made of rice, so rice flour seemed to be a logical ingredient, consonant with the dish, common in India and cheap.
Little by little, we succeeded to design an easy recipe, to cook small cubes made of rice flour and water: the VitaCubes. The idea was also to be flexible to every VitaCube maker means, therefore the recipe is not very strict and can be adapted to what the people have available.
Now that we have found a convenient distribution mean, we need to be sure that it keeps our strains alive and to know how long it can be stored.
We conducted several survival test on the VitaCubes, using S. cerevisiae and L. lactis. We observed that the yeast can be stored more than one week in a VitaCube. For the bacteria L. lactis, we found that after 4 days of drying, the survival rate started to be very very low. Nevertheless, there are still around 10^5 cells in a VitaCube after one week, which is far enough to make a culture from it.
European and Indian Regulations
In order to evaluate the feasibility of our product’s implementation, we researched the european and indian regulations concerning the production and distribution of genetically engineered micro-organisms.
We found that the EU directive 90/219/EEC of the European Economic Community relative to the contained use of GMO would allow production of this product within the european market. This directive is enforced in each of the EU member’s national regulations.
In India, the FSSAI (Food Safety and Standard Authority in India) told us that the safety is needed to be established in order for our product to be authorized. Both Dr. A. K. Sharma from the FSSAI and Dr. Sunita Grover the Dairy Microbiology Division at the National Dairy Research Institute advised us to chose micro-organisms that were already present in the fermented foods we were targeting, which is what we did. These organisms are all in Risk Group 1 (Unlikely to cause human disease.), and have the GRAS status (Generally Considered As Safe).
We also found out that the Indian law currently doesn’t allow GM microorganisms because of the use of antibiotic markers that makes them unsafe to eat. So we'll have to remove all antibiotic markers from our strains.
According to Samir K. Brahmachari, former director of the Council of Scientific and Industrial Research in India, our product has a greater chance to be authorized if there is no more live bacteria in the final form of the dish, that is actually eaten. We checked this affirmation and found out this regulation:
‘...food stuffs...derived from Living Modified Organisms where the end product is NOT a Living Modified Organism are exempted from mandatory approval of the Genetic Engineering Approval Committee.’
Food Safety and Standards Authority of India (FSSAI)
Notification no. S. O. 1519(E) dated 23-8- 2007 in the Gazette of India
Since the idli is steamed for 10 to 15 minutes before it is eaten, we made an experiment to assess the presence of yeasts and bacteria in the steamed idli, and found out that the steaming process effectively sterilizes the idli and eliminates the microbes present in the batter. On the photo above, we can see than nothing grew in the plates inoculated with steamed idli batter, in the 5 different media we tested - which indicates that all the micro-organisms present in the batter were killed during the steaming process.