A better understanding of microgravity, vacuum, and space missions was key to the success of our project, as microorganisms and their functions are extensively impacted by these conditions. The precarious nature of space and how it would impact the material and structure of our 3D printed bioreactor was difficult to comprehend. We thus needed expert input to find ways of sustainable production of edible quality protein while incorporating our bioreactor without having much impact on the payload and its corresponding cost for space missions.

One challenge we faced was not knowing how astronaut waste is currently processed, which made it difficult to assess the relevance of our focus on nutrition and recycling. We also questioned whether melanin films and genetic modifications like Dsup and Rad51 could adequately counter space radiation, while evaluating the feasibility of cultivating protein from yeast in space and the potential long-term impact of our project on future missions.

A major challenge we faced was understanding the radiation environment in space, particularly the types and intensities encountered beyond Earth’s atmosphere. Additionally, we needed information on the economic feasibility of deploying a payload, as costs vary significantly across space agencies. These aspects are crucial in assessing the long-term applicability of our astronaut nutrition project.

Rohan explained the three major orbital zones—LEO, MEO, and GEO—and highlighted that radiation intensity increases with altitude. He detailed the radiation sources astronauts face: ultraviolet (UV) radiation, cosmic rays, X-rays, solar flares, and high-energy protons and ions of hydrogen and helium. He also described the Van Allen radiation belts and their impact on spacecraft and biological systems.

On payload costs, he outlined comparative figures: ISRO’s GSLV (~$7,000–8,000 per kg), SpaceX ($2,000–4,000 per kg, with ambitions of reaching ~$1,000), and ULA/Firefly (> $10,000 per kg). He emphasized that while costs are decreasing, higher orbit launches remain expensive. for our project, he suggested testing the concept using a CubeSat with a miniaturized bioreactor, ensuring that it is chemically inert and non-reactive. He advised determining astronaut protein requirements, calculating urea-to-protein conversion efficiency, and designing for microgravity by using an injection mechanism where nutrient media spheres could be broken down and dispersed.

Based on his advice, we began exploring CubeSat-based testing as a practical and low-cost proof-of-concept pathway. We evaluated the design of inert and non-reactive materials for the bioreactor to ensure safety in microgravity. Our team also initiated calculations for astronaut protein requirements and urea conversion ratios to validate the nutritional feasibility of our system. Inspired by his microgravity solution, we considered incorporating injection mechanisms into our reactor design for improved nutrient dispersion and functionality in space conditions.

Astronauts are central to our mission, as their safety, comfort, and nutritional needs shape every aspect of our project. Dr. Suresh Naik had advised us to thoroughly understand the workability of our project to create a space worthy system. To ensure our system is practical for long-duration space missions, we needed the help of an astronaut to understand current food logistics, microbial safety protocols, and operational feasibility aboard the ISS.

Through our interaction with Michael, we realized the need for strict containment protocols and PPE when working with microorganisms. He confirmed that astronauts could manage working with complex systems with proper training. To enhance usability, he suggested developing an easily consumable ‘SCP liquid’ tailored for onboard routines. While taste wasn’t a major concern, he emphasized that the product should be at least palatable. Most importantly, he urged us to quantify the advantages of our system and clearly demonstrate its operational feasibility and nutritional impact for long-duration space missions.

We started defining key features of our project, such as protein yield per gram of urea, cost per meal, and shelf-life extension. Our bioreactor is being designed with containment layers and astronaut-safe interfaces to minimize microbial exposure and ensure operational safety. Along with this, we are also exploring SCP formulations to achieve taste neutrality, while also investigating processing methods that allow SCPs to be consumed in liquid form, thus making them more compatible with astronaut routines and onboard constraints.

Our project involves genetically modified yeast engineered to produce edible single-cell proteins (SCPs) within a space-operable bioreactor. This raises critical biosafety concerns, including the potential release of GMOs, risks of antibiotic resistance, and the implications of accidental ingestion by astronauts. Additionally, we needed to understand the risks posed by bioreactor materials, and the disposal of biological waste in space. The possibility of engineering radiation-resistant yeast further complicated biosafety and sterilization protocols, making it necessary for us to have an expert opinion.

For our project on nutrient production for space, we faced challenges in maintaining sterility and hygiene during food production. Ensuring that astronaut food is produced in a controlled, contamination-free environment is critical for both safety and nutritional reliability. We needed insights on industrial-scale sanitation and quality control protocols to design our processes effectively.

Our team faced uncertainty in systematically assessing the risks associated with our project. We needed clarity on whether our organism or its modifications could present biosecurity concerns. We wanted to understand the potential consequences of a space leak and whether toxicity and palatability testing would be sufficient to demonstrate safety to iGEM judges. Additional concerns included how to manage sterilization and containment in space, the potential for microbial mutations under radiation, and whether risks identified should be explicitly mentioned in our iGEM wiki.

After attending the Dual Use Research Workshop held at the All India iGEM Meet, organised by iGEM Ambassadors, we realised the need to evaluate our project from a broader perspective. In doing so, we realised the unintended potential of our project for dual-use concerns, particularly given our work with radiation-resistant microorganisms. We thus needed Christopher's help to clarify our suspicions and explore ways to responsibly identify and mitigate any shortcomings in our project.

Christopher commended our efforts to identify and eliminate the potential dual-use of our project. He assured us that teams that proactively highlight such risks in their project are not penalized in medal qualifications but are rather appreciated by the judges. He advised us to further explore ways of preventing such adverse impacts and to create a Risk Analysis for our genetically modified organism and the bioreactor.

As advised by Christopher, we have begun reading about the overexpression of DSUP in prokaryotes, as historically, prokaryotes have been utilized for military purposes. We are actively assessing the potential risk of using a bioreactor and evaluating options to mitigate any adverse impacts that may arise from its use.

The main challenge was establishing contact with such a high-level policy-making body. Since DBT’s formal iBEC program ended in 2020, there was no existing channel for iGEM teams to reach out. Coordinating pan-India representation and finding an effective approach to DBT posed an additional challenge.

To address this, our team independently gathered consent from all registered Indian iGEM teams and contacted iGEM HQ, obtaining an official confirmation of the Indian student delegation. IIT Madras later joined the effort, with support from its Dean of Students Office, which formally forwarded our request for dialogue to DBT on behalf of all Indian teams. While we did not receive a meeting, the communication was acknowledged and the delegation letter was successfully forwarded to DBT.

Although no direct consultation took place, this initiative demonstrated the feasibility of coordinating pan-India efforts and re-establishing contact with DBT. It highlighted the importance of building long-term student-policy engagement channels, which can inform future interactions on synthetic biology policy, funding, and representation. Importantly, the attempt itself became a valuable iHP activity, as it showcased our proactive approach in reaching out to the highest level of policy makers relevant to our field.

Community response and social impact are of paramount importance for our project. We wanted to further reach out and spread the idea of synthetic biology in space and biosafety among the masses, for which we were looking for media opportunities and individuals to connect with. We also wanted to create a biosafety manual so as to ensure that our project aligned with the UN’s Sustainable Development Goals.

We are actively collaborating with Ruchira to amplify the impact of our project. With her guidance, we have identified biosafety officers and legal experts and have reached out to Manaswar Space, astronauts, and relevant officials. These efforts aim to align our project with the Sustainable Development Goals.

On advice of Jasmin Ponda, iGEM - ABOA, we began looking for a structuring a business plan for our project however we encountered difficulty in creating a robust business model and commercialisation pathway for our project. Further, we also needed her help in understanding ways to improve our project presentation, to possible investors and agencies.

Shreya advised us to pitch our product exclusively for space missions. Positioning it for astronauts gives us a unique edge and avoids being seen as just another protein powder company. She also advised us to gather astronaut responses to our project. We discussed two commercialization routes: selling the strain or selling the bioreactor system. The latter allows for modular, on-demand production and aligns better with investor expectations. She advised us to include a timeline showing when the product will be ready for terrestrial pilots, when it will be pitched to space companies, and when validation milestones will be achieved.

We’ve shifted our focus to astronaut nutrition, making our project stand out from regular protein solutions. We started building a basic financial model to show how our system could save costs compared to sending food to space. Our business plan now includes a subscription-based model to ensure long-term use and reliability. We are looking into mapping out a timeline for the strategic release of our products to investors. We also held a meeting with astronauts to gather their feedback and identify key areas for improvement in making our system space-ready. IP as a source of income and revenue.

Our team faced two main challenges: (1) developing a concrete entrepreneurship plan for the iGEM deliverables, which includes market analysis and audience mapping, and (2) identifying practical ways to scale our 3D-printed bioreactor and protein flakes beyond purely space-centric applications. Since the commercial space food market is very limited, we needed guidance on how to broaden our impact while still retaining our project’s originality.

Mr. Tibrewal encouraged us to think beyond space technology, noting that entrepreneurship in such a narrow sector poses difficulties due to the small market size and high entry barriers. While addressing astronaut nutrition is valuable, he stressed the importance of positioning our solution in broader markets, such as tackling food security and accessibility in India, where catering to a billion people could be transformative. He emphasized that entrepreneurship requires both depth in a core domain and adaptability across sectors, and engaged with our ideas for a 3D-printed bioreactor and protein flakes. Most importantly, he highlighted the pathway from ideation to market launch, stressing the need to analyze problem scope, identify target audiences, and avoid restricting applications to a single domain. He also shared a presentation on student entrepreneurship, offering insights into ideation, problem validation, and balancing scientific innovation with business viability.

Based on his feedback, we began reframing our project pitch to highlight its potential in addressing global food challenges in addition to astronaut nutrition. We also expanded the entrepreneurship section of our documentation to include broader market analyses, considering protein supplementation for terrestrial applications alongside space missions. His insights guided us to view our bioreactor not just as a space technology but as a platform for sustainable food production at scale. Furthermore, his emphasis on audience and segment diversification has ensured that our project remains both scientifically innovative and commercially viable.

Our team faced challenges in adding flavors to protein formulations while maintaining both taste and chemical stability. Previous iHP consultations with astronauts emphasized that taste is a critical factor in long-duration space diets, as unpalatable food can negatively affect nutrition intake and morale. Furthermore, incorporating natural flavors rather than artificial ones would make the process more sustainable, offering a major advantage for space applications and aligning with our goal of safe, clean, and efficient nutrient production.

We consulted Jalinder Shinde, who is in charge of protein powder production at The Whole Truth Foods. He provided valuable insights into how the company uses natural ingredients such as Monk fruit, Date sugar, Cashew butter, and other plant-based products to enhance taste without relying on artificial additives. He explained that these ingredients are carefully tested in their lab to determine the correct proportions needed for a balanced and stable flavor profile, ensuring compatibility with the overall product while maintaining sensory appeal.

This consultation helped us understand the complexities of flavor formulation, particularly the need for precise measurement, stability testing, and sensory evaluation when using natural ingredients. These insights guided our approach to developing protein formulations suitable for astronauts, ensuring that they are nutritious, palatable, and sustainably flavored, while maintaining chemical stability and compatibility with the rest of the formulation.

Our primary concerns were defining the business scope, evaluating the economic feasibility of both space and terrestrial applications, and addressing ethical and policy challenges related to GMOs and public acceptance. We had questions about the market feasibility for astronaut-focused protein systems.

Jasmin suggested starting with narrow, high-need markets like space missions, rather than attempting to tackle global food scarcity immediately. Conducting ethics and policy consultations to anticipate regulatory hurdles and public acceptance issues. Building a clear business strategy that balances scientific innovation with feasibility, scalability, and impact.

We will redefine our business model to prioritize astronaut nutrition and other specialized applications while keeping long-term sustainability and community impact in mind. Ethical considerations and regulatory pathways will be explicitly included in our project narrative to address concerns about GMO-based products.

Our team needed clarity on two major aspects of space nutrition: (1) How astronauts’ nutritional needs differ from those on Earth, and (2) whether SCP production from urea could realistically address critical deficiencies or reduce reliance on costly resupply missions. Literature indicated persistent issues such as muscle loss (sarcopenia), energy deficits, and limited menu options in space, but the data were fragmented. We required expert input to position our project within these nutritional challenges.

Scaling up our project from the confines of the lab to the real-world platform was our next step. To ensure sustainable and successful scale-up, we wanted to understand the variables that would impact our project's performance at the industry level. We also wanted to ensure that our plan for upscaling is aligned with the industry-accepted norms in terms of protein purity and estimating process efficiency.

We identified key areas to refine in our project, starting with understanding the exact protein extraction mechanisms and ensuring the nutritional profile aligns with astronaut dietary needs. He emphasized post-production processes such as protein isolation, selective precipitation for by-product separation, and waste recycling. A crucial next step involves developing a stable and consumable protein formulation tailored for space missions.

After the consultation, we focused on protein extraction and formulation techniques highlighted by the expert. We studied selective precipitation methods to separate by-products efficiently, explored wave-back reactor designs to improve process stability at scale, and refined nutritional profiling to ensure the protein meets astronaut dietary requirements. We also researched strategies to make the protein stable, palatable, and safe for space missions, helping us align our scale-up plan with industry standards and ensure a robust, space-ready final product.

Our team needed input on optimizing the cultivation and harvesting of yeast for single-cell protein (SCP) production. Specifically, we wanted to know regarding nutrient requirements, the stage at which yeast should be harvested to maximize protein content, methods for protein recovery, and approaches to minimize off-flavors. Furthermore, we sought advice on how these processes could be adapted for anaerobic fermentation in space environments, where conditions such as pressure, temperature, and gas balance differ significantly from Earth.

Dr. Annapure highlighted the essential nutrients for yeast growth, including carbon, calcium, phosphorus, sodium, sulfur, and other micronutrients. He emphasized harvesting yeast at the junction of the log and stationary phase, when nitrogen content is highest and before the production of secondary metabolites. For protein recovery, he recommended mechanically drying the yeast, lysing the cells, and isolating proteins through precipitation methods such as salt precipitation or HPLC. Since SCP often imparts off-flavors, he stressed the importance of isolating pure protein for better acceptability.

He also noted the importance of monitoring O₂, CO₂, pH, temperature, and pressure, while advising us to specifically check the temperature range maintained on the International Space Station . To separate biomass from by-products such as alcohol and CO₂, he suggested filtration techniques. Additionally, he cautioned that certain nitrogen sources could damage yeast and should be carefully selected. For practical applications, he suggested that SCP could be incorporated into normal food items such as cheese or butter, making it more palatable for astronauts.

Based on his inputs, we refined our wet lab design to focus on harvesting yeast at the optimal growth stage to maximize protein yield. We are now incorporating downstream processes like mechanical drying, cell lysis, and protein precipitation into our workflow, ensuring that flavor issues associated with SCP are minimized. Furthermore, we are considering anaerobic fermentation setups suitable for space conditions, with careful monitoring of environmental factors such as pH, gas balance, and pressure. His suggestion to integrate SCP into familiar food products has also been adopted into our application strategy for astronaut diets.

While developing our project on nutrient-rich protein production for space applications, our team was uncertain about the accuracy and industrial validity of our protein estimation method. We specifically wanted to confirm whether the Kjeldahl method we used in our experiments was consistent with food industry standards and whether the nitrogen-to-protein conversion factor (6.25) would remain applicable under space-like conditions.

To address these questions, we consulted Sameeshka Shetty from the Quality Lab team of a reputed health food company. She confirmed that the Kjeldahl method is indeed the industry-standard technique for protein estimation. The process involves digesting the sample, measuring its nitrogen content, and using a conversion factor (typically 6.25) to calculate the protein percentage. She further clarified that this factor would not vary significantly in space, validating our assumption. Additionally, her insights on the company’s use of natural ingredients and preservative-free formulations highlighted practical approaches to maintaining nutritional purity and safety.

This consultation provided strong validation for our experimental approach. It confirmed that our protein estimation technique was scientifically sound and aligned with industry practices, strengthening the credibility of our data. Furthermore, the emphasis on natural and transparent processing methods inspired us to incorporate similar principles into our Single-Cell Protein (SCP) production framework—ensuring that our system for space nutrition remains safe, sustainable, and free from artificial additives.

We sought guidance on how to frame our project clearly and effectively for both technical and public audiences, aiming to present it under a single cohesive framework. In the process, we discovered that there is limited literature on the effects of microgravity on yeast metabolism and on how different types of radiation impact microorganisms, highlighting gaps that our project may help address.

Our challenge was to identify a feasible and safe way to test the radiation resistance of our melanin-coated yeast cultures. While Earth-based lab experiments gave us preliminary findings, they could not fully replicate the microgravity and radiation stress of near space conditions. To validate our results, we needed exposure to space-like radiation. However, with limited options available, we needed expert guidance to determine the most suitable method.

During our discussion, Dr. Ninan outlined two experimental approaches. The first was the TIFR high-altitude balloon facility, which can carry biological samples up to 30–40 km, exposing them to cosmic radiation and UV at near-space levels. For safety, the samples would contain baker’s yeast (Saccharomyces cerevisiae) instead of our engineered strains, coated with melanin to test its protective efficiency. The second option was the LINAC (Linear Accelerator) facility, which simulates cosmic rays through the use of particle beam irradiation. However, this method involves direct bombardment of particles and localized heating, creating a risk of damaging the yeast cultures. After evaluation, we chose the balloon facility as the safer and more effective testing environment. We also discussed practical considerations, including altitude, flight duration, and monitoring of samples.

Based on Dr. Ninan’s guidance, we finalized the balloon experiment using melanin-coated baker’s yeast, a safe and non-pathogenic model widely used in space biology. The test has been confirmed, but will be conducted after the WikiFreeze. Once executed, this experiment will allow us to assess the protective efficiency of melanin against space-like radiation.

Our main challenges were deciding how to analyze the amino acid content of our protein product, determining whether intracellular protein production by yeast will require extracellular secretion for easier recovery, and establishing pathways for toxicity and palatability testing. We also faced limitations in testing radiation resistance directly in the wet lab due to equipment and safety constraints. On the technical side, designing a closed-loop bioreactor system, including oxygen recirculation, sizing considerations, and downstream processing, posed additional complexity. Questions about technical feasibility for both space and terrestrial applications also arose.

Our dry lab needed some guidance in modelling population balances for yeast in the bio-reactor, incorporating Damage Suppressing Protein (DSUP), and validating its behaviour and performance experimentally. Additionally, we needed advice on how to present our modeling work, document our progress, and align our entrepreneurship documentation with iGEM standards.

Our team needed expert guidance on the effects of microgravity on our system intended for use aboard the International Space Station and space missions. Since the laws of physics in microgravity differ significantly from that on Earth and the available literature is limited, we sought his advice to address these uncertainties.

In our discussion with Mr. Pariccha, we learned that our system would need to rely on diffusion rather than sedimentation or buoyancy. He recommended the use of peristaltic pumps for water circulation, noting their durability and the fact that ISS maintains constant temperature and pressure, so vacuum conditions are not a concern. To minimize bubble formation, he suggested ensuring continuous flow, using gas filters, or incorporating hermetic seals. For radiation protection of the bioreactor, he advised supplementing our genetically engineered melanin with ABS coating. He also recommended epoxy sealants for biosafety and testing the system in thermo-vacuum chambers to simulate space conditions. He suggested using 3D printers in space would be more better as the risk of the reactor being damaged during launch is indeed.

Following his suggestions, we adopted the use of peristaltic pumps in our design and decided to combine ABS with melanin for enhanced radiation protection of the bioreactor. We are now designing our reactor to account for all necessary safety parameters based on his recommendations.