MikuyTec

Mikuytec proposes large-scale recombinant production of antifungal and antioxidant peptides for their use in a coating applied to fruits to slow down spoilage and extend their shelf life.

MikuyTec’s main objective is the extension of the shelf life of fruits and vegetables to reduce product loss during transportation and distribution stages. We propose a large-scale recombinant production of antifungal and antioxidant peptides for application in a coating applied to crops. To address that, we designed a scalable bacteria, E. coli BL21, to express two genetic circuits or cassettes through synthetic biology. The first one encodes 3 antifungal peptides (Rs-AFP2, PAF26 and PAFP-S) to which a specific domain (CBD) is added to improve the specificity of action towards fungi. While, the second one synthesizes the antioxidants peptides (VS14 and NOP-1) to inhibit the tyrosinase enzyme, responsible for the fruit browning. It should be noted that the peptides of both cassettes were selected from a library of chimeric proteins made with different analyzes and bioinformatic modeling. As well as the verification of its large-scale production in bioreactors with mathematical models.

Finally, the peptides will be extracted from the bioreactor through centrifugation and lyophilized into powder for delivery to fruit and vegetable distribution companies. This powder can later be dissolved in water so that the peptides regain their biochemical activity and can be safely sprayed onto the crops.

Of the nearly 26.6 tons of fruits and vegetables produced each year in Peru, around 12.8 million never reach markets or homes. When analyzing the supply chain of food, we realized that an average of 47.6% of the country’s annual food production is lost, which places us among the highest rates of food waste in Latin America. We identified that this degradation process occurs mainly due to fruit oxidation and the opportunistic fungi during the transportation process. This problem represents a challenge for environmental sustainability and also affects our population in different ways. Farmers and distribution companies receive less income by not taking advantage of all the production. Likewise, the precarious state of the supply chain in remote areas of the country exacerbates the already existing problem of malnutrition. To help mitigate this problem, various technologies are used to help keep food fresher for longer, such as chemical preservatives, which can have adverse health effects when consumed in high concentrations and many times can be washed into nearby water bodies harming local wildlife. An alternative is a constant refrigeration, however, this tends to be expensive and difficult to access, especially in remote areas of the country. Taking all this into account, MikuyTec seeks to aid the agricultural and agro-export sectors, two of the largest economic sectors in the country, since it could greatly benefit from biotechnology.

MikuyTec will help mainly fruit and vegetable distributors, farmers and end consumers. It will increase the shelf life of the fruit so that they will have much more time to sell and economic post-harvest losses will decrease, it will combat the deterioration of fruit in the transport stages so that much more raw material will reach the market at more affordable prices for the final consumer and finally it will increase food safety by decreasing the likelihood of diseases caused by fungal toxins and access to quality food in remote communities.

We decided to investigate the needs of our target population. In the first place, we use methodologies such as Design Thinking to understand the problem they face and thus empathize with their needs and propose a solution that best suits their needs. We visited the main local markets in Lima, Peru and conducted interviews and surveys of fruit distributors. This was essential to propose our solution, to develop it and to test it. By having the design of our solution using synthetic biology, we were able to expose the idea to local markets through an infographic summary, with this we obtained insights, as a result, the public liked the idea of the project and how it attacks the problem of fruit decomposition at the time of distribution. Finally, we are holding informative workshops and talks on what the project is about in our scientific community and soon we will hold workshops for the general public so in that way they can learn more about the science behind our proposal, Synthetic Biology. and how it works.

We are transitioning from Concept to Prototype, but mainly, in the second stage. This is because we have already developed the product with computational tools. We have completed the design of the 2 genetic circuits for the recombinant production of antioxidant and antifungal peptides, respectively. Likewise, we have the plasmid and the experimental protocols ready for the genetic transformation of the bacteria that will produce our product. 

Furthermore we made the testing stage at a technical and social level. For the selection of the peptides, we generated a library of chimeric proteins with the selected genetic parts to conduct a structural analysis and modelling of its potential antifungal activity through several bioinformatics tools. And we developed math models to predict the bacteria growth at bioreactors and fungal growth inhibition. On the other hand, we tested this product with 12 farmers from the community of Cañete, Lima (Peru) and 2 agro-export companies (CBC company and AGJ CORPORATION SAC). Which helped us to improve the presentation of the product, the biotechnological explanation before an audience that does not have as much knowledge about the subject and to continue validating and learning more about the problem.

The core technology that we use for the development of our project is synthetic biology, an interdisciplinary area that involves the application of engineering principles to biology. It aims at the redesign and fabrication of biological components and systems, like the one we propose in our proposal.

We plan to make use of genetic circuits in a bacterial mode to express recombinant proteins, making use of bioprocesses and bioreactors to scale production of these peptidos to higher levels.

We aim to offer our product to 10% of the total number of agro-export companies in Peru (1986 companies) in the next 2 years. After that, we want to increase that percentage in our country to 50%. In 10 years, MikuyTec will be expanded in the global market of all the continents. These are the numbers of agro-export companies established in each of them:

South America: 1874+

North America: 6858+

EU, Africa & ME: 38027+

Asia & Oceania: 26438+

It should be noted that each of these companies will have a great impact on the farmers who produce the products and an undetermined number of people that buy those fruits and vegetables at the market. We still can not know the exact number because it depends on the statistics handled by each company.

In order to achieve our main objective of extending the shelf life of fruits and vegetables, we proposed a plan in the different committees that we have. In the first place, we are going to iterate and improve the technological part of our product through consultancies with the company Ginkgo Bioworks and workshops that we are generating among ourselves. On the business side, we are applying to different entrepreneurship or bio-entrepreneurship programs to refine our business model. On the legal side, we are making a new call to bring together professionals to help us introduce the product and startup in all of Peru's legislation. In the financial part, we are forming an alliance with the UTEC to contact agro-export companies interested in investing in projects like ours. In the part of the human practice, we are developing and planning workshops for students with the aim of promoting synthetic biology and receiving greater support for biotechnology. Finally, with all these steps and the support of our sponsors from the iGEM design league competition, we plan to have our first tested prototype by the end of this year, as a result of future participation in the global iGEM competition.

We aim to measure the impact of our project through two indicators in the next year. One of them is the effective amount of extended shelf life our product provides and the new time window it provides producers for placing their products. For this purpose, we plan to conduct experiments aimed to determine the amount of days our product can extend fruit’s shelf life and to evaluate the use of other elements or changes in our product that could improve this amount. On the other hand, outside the experimental results that we will obtain, we consider it important to determine how it benefits the use of our product to our consumers. We plan to conduct surveys to know how much extra time to place and sell their products they have achieved and if this is worthwhile. Also we plan to monitor several batches applied with our product to determine the effectiveness of our product on preventing fruit spoilage. Furtheremore, we we are measuring our progress since the fulfillment of 3 SDGs: #2 End hunger, achieve food security and improve nutrition and promote sustainable agriculture, #12 Responsible consumption and production, and #13 Take urgent action to combat climate change and its impacts.

First of all is the visa procedure to be able to travel to North America and that Ginkgo can advise us in a more practical and personal way. We have problems to be able to redeem this prize that we obtained based on our efforts in the Igem Design League competition. Other barriers are the altered prices in the market because of Covid and the political fragility in which our country currently finds itself. Another complication would be that we do not have enough money to be able to continue with the next stages, we are still trying to get funding. Many of us are still students and have financial shortcomings, but the will to go ahead is strong. 

We are a team made up of students from different parts of Peru. Each one in their own region found a common problem which is fruit spoilage due to fungus. We are an interdisciplinary team of bioengineering, biology, agroindustrial engineering, genetics, and biotechnology students. This allowed us to study the problem from different points of view. With all this background we create the design of a biological system with synthetic biology. Now we are divided into different work areas such as Web Lab, Dry Lab, Entrepreneurship and Human Practices to ensure correct implementation of the project. During this current period, we have contact with agro-export companies that are our main users.

Last year we obtained first place in the iGEM Design League competition, the most important Synthetic Biology competition in Latin America, and as a result of this, we obtained some awards. Our main sponsor is Ginkgo Bioworks, who will finance our trip and stay in Boston to visit their facilities. Likewise, in the following month we will begin workshops with this company with the aim of refining the product and preparing ourselves for the elaboration of the first prototype and our training as a startup. On the other hand, IDT DNA will provide us with the sponsorship for the synthesis of plasmids and all genetic material. Finally, we are in the process of establishing an alliance with the Universidad de Ingeniería y Tecnología to contact several companies in the field that can finance the implementation of the product.