BFH Frankfurt
We attended the BFH-Euro Meet-Up from the 23rd to the 25th of May. This was our first international iGEM conference, where we got a chance to connect with iGEM teams from all over Europe. The event was filled with inspiring pep talks, helpful workshops, and many opportunities for networking, socialising, and exchanging experiences with other teams.
During the event, we had the chance to present our project in our very first poster session and we were honoured to receive the Team Booth Award. This recognition was a great motivational boost for our team to continue our hard work on the project.
We also received valuable feedback from participants and experts. One suggestion from an iGEM Judge was to verify the safety of Rhodococcus for food-grade and medical-grade production and to check whether it produces any toxins that could contaminate the product. Although we were not aware of any problematic side producst,we searched the literature to ensure thoroughness. We found that Rhodococcus has already been used to produce pharmaceuticals [1] and food-related products such as carotenoids [2], confirming its suitability for our application.
Another participant proposed using DFO for wastewater treatments and recommended contacting Dr. Rohan Jain, an expert in this field. We gained practical insights into effective poster design and presentation.This meet-up was a great opportunity for our team, helping us to build connections and improve on both scientific and presentation aspects of our project.
iGEM Meet-Up Düsseldorf
On the 5th of July, Marvin attended an iGEM Meet-Up in Düsseldorf. The event featured inspiring presentations and offered many opportunities for discussion and idea exchange among teams. We had the opportunity to compare our project development with other teams and receive constructive feedback that we integrated integrated into the subsequent stages of our work.
During our presentation, the audience raised insightful questions regarding our choice of Rhodococcus as a host organism. One suggestion was to consider using Corynebacterium, which is already widely used in industry for large-scale amino acid production, including lysine, which serves as the starting point of the DFO biosynthesis pathway. Another point raised was the potential toxicity of DFO to Rhodococcus, as it might chelate iron, resulting in iron starvation. As a result of this feedback, we decided to perform growth curve experiments that showed that DFO did not have a significant effect on the growth behaviour of Rhodococcus, suggesting that Rhodococcus is either tolerant to DFO or may be able to utilize it for iron acquisition.
The discussions provided valuable perspectives that helped us better understand the strengths and potential challenges of our chosen system: for example, that Rhodococcus might deplete its lysine pool during DFO production, requiring lysine supplementation in the growth medium. While Corynebacterium would address this concern due to its natural lysine overproduction capacity, we chose to continue with Rhodococcus given our project timeline and established protocols for this organism.
Interview Prof. Julia Bandow
On 23 July, we interviewed Prof. Dr. Julia Bandow, a microbiology professor at Ruhr-University Bochum with direct experience in DFO production and analysis. Prof. Bandow previously worked for Pfizer Global Research and Development and has published on desferrioxamine biosynthesis. We sought her expertise on purification methods, analytical techniques, and regulatory considerations for our Rhodococcus-based production system. She emphasized that she appreciates our approach, but also pointed out that we need to keep GMO regulations in mind when incorporating genes from other organisms into ours.
Why did we choose to talk to Prof. Bandow?
- Published research on desferrioxamine biosynthesis in Streptomyces
- Expertise in mass spectrometry analysis and metabolomics
- Experience in pharmaceutical research and development, worked for Pfizer Global Research and Development (was awarded the Pfizer Clinical Research and Development Award in 2006)
- Familiarity with European GMO and pharmaceutical regulations
Main topics during the interview:
- Analytical methods such as MS-analysis for DFO detection and quantification
- Iron removal strategies for DFO purification
- Regulatory considerations for genetically modified production strains
- Organism selection and metabolic considerations such as DFO uptake and export and how it could interfere with our project
- Downstream processing and formulation requirements
What did we learn?
- Analytical approach: Standard mass spectrometry is unsuitable for iron-loaded DFO-samples. Prof. Bandow recommended ICP-MS (Inductively Coupled Plasma Mass Spectrometry) and offered collaboration for analysis using their equipment.
- Purification strategy: Magnetic bead-based iron removal could simplify downstream processing by capturing iron-DFO complexes before releasing iron-free DFO.
- Organism considerations: Prof. Bandow noted that fast-growing organisms like Rhodococcus possess iron storage systems that may sequester iron, potentially competing with DFO production. She suggested Streptomyces as an alternative. However, we maintained our focus on Rhodococcus due to its established genetic tools, shorter production times, and ability to utilize diverse substrates.
- GMO compliance: Prof. Bandow also reminded us that we should check for compliance with European GMO regulations.
- Application-specific formulation: For pharmaceutical applications, DFO must be formulated for specific delivery methods (e.g., injectable solutions require particular solubility and stability profiles).
Impact on our project: Prof. Bandow's insights guided our analytical strategy and reinforced the importance of regulatory compliance. While she raised valid concerns about iron storage in Rhodococcus, our growth curve experiments (conducted following the Düsseldorf meetup feedback) demonstrated that DFO does not significantly inhibit Rhodococcus growth.
Junior Jam Münster
As part of our iGEM journey, we attended the JuniorJam Meetup in Münster from the 25th to 27th of July. There we had the opportunity to connect with teams from across Europe, exchange ideas, and present our project. This event allowed us to build new connections and strengthen existing ones. Also it offered valuable feedback from both industry experts and iGEM judges, this feedback offered us new ideas about and prepared us for the Grand Jamboree in Paris
During our presentation, the judges raised several critical questions about our project. The discussion highlighted that we can improve on our presentation skills and on clearer communication of our scientific reasoning. They also encouraged us to better emphasise the novelty and advantages of our approach.
During the meet-up we had the opportunity to receive valuable insights from industry experts, such as potential use of alternative carbon sources. We had initially considered using glycerol as a cheaper alternative to glucose, with the idea of preserving glucose for human food use. However, the expert explained that on an industrial scale, glucose is actually cheaper than glycerol and that carbon sources contribute only marginally to the overall cost of pharmaceutical production. Additionally, we discussed regulatory considerations and the importance of human practices in strengthening the societal relevance of our project.
This Meet-up was highly valuable, both for networking and for the constructive feedback we received. This experience contributed to broader understanding of the nuances of the industry field and made us better prepared for the Grand Jamboree.
Interview Dr. Arno Cordes
During our project we interviewed Dr. Arno Cordes, CEO of ASA Spezialenzyme and specialist in biocatalyst production. His company produces enzymes for industrial applications, including wastewater treatment. Dr. Cordes discussed DFO synthesis methods employed by his company and emphasized that DFO yields of 10-100 g/L are necessary to achieve commercial viability. He offered to compare our Rhodococcus production system with their established methods, which would provide valuable benchmarking data for our production process. He also recommended we contact Dr. Rohan Jain at the Helmholtz-Institut Dresden for additional expertise.
Why did we choose to talk to Prof. Cordes?
- Industry expert with direct experience in commercial DFO production
- Specialist in industrial-scale enzyme and biocatalyst manufacturing
- Insights into economic viability and optimization strategies
- Aspects of DFO-production, how important is reducing the costs of C-sources?
- Would Rhodococcus be a viable option for industry, as a DFO-production platform?
- How DFO yield ould be enhanced
- DFO variants: DFO-E (desferrioxamine E) production methods are similar to DFO-B, suggesting our Rhodococcus expression system could be adapted to produce both siderophore variants, broadening commercial applications
- Industrial applications: Beyond medical chelation therapy, DFO-B is used for recovering rare earth metals and rust removal, though current production costs limit these applications
- Cost optimization priorities: Nitrogen sources are approximately five times more expensive than carbon sources, making nitrogen source optimization an attractive starting point for reducing production costs.
- Yield improvement strategies: DFO yields could be enhanced by blocking competing metabolic pathways that divert biosynthetic precursors away from siderophore production.
- Commercial viability threshold: Achieving yields of 10-100 g/L is essential for industrial-scale production to be economically competitive.
Main topics during the interview:
What did we learn?
Impact on our project: Dr. Cordes insights reinforced our focus on Rhodococcus as a production platform and highlighted the importance of metabolic pathway engineering and nitrogen source optimization for achieving commercially viable yields.
Interview Dr. Rohan Jain
During our Human Practices work, we contacted Dr. Rohan Jain, Group Leader at HIF, Helmholtz-Zentrum Dresden-Rossendorf. In our conversation, Dr. Jain emphasized that while Desferrioxamine has pharmaceutical relevance, environmental applications such as wastewater treatment may be a more suitable direction for our work. He confirmed that if we would be able to produce large amounts of Desferrioxamine within a short period of time, this would align well with the growing demand for new technologies in environmental management. Importantly, he also pointed out that, unlike in the pharmaceutical sector, the purity of Desferrioxamine is less critical for wastewater treatment. This makes the goal of producing large-scale quantities more realistic and attainable for our project.
Beyond these technical considerations, Dr. Jain underlined that the most decisive factors for industrial application are cost and quantity. His advice broadened our perspective, instead of focusing solely on pharmaceutical use, we could look into strategies that lower production costs and increase yield.
Why did we choose to talk to Prof. Jain?
Dr. Rohan Jain is Group Leader at HIF, Helmholtz-Zentrum Dresden-Rossendorf. He is an experienced Environmental Technologist with a history of working in the wastewater and recycling industry. He authored a paper on Recovery of gallium from wafer fabrication industry wastewaters by Desferrioxamine B and E using reversed-phase chromatography approach. Dr. Jain has extensive experience with siderophores, having participated in projects such as:
- Biotechnologische Verfahren zur Rückgewinnung von Germanium, Indium und Kupfer aus kupferhaltigen Industriestäuben (with Dr. Katrin Pollmann)
- SideroFlot Phase 2: Siderophore als mikrobielle Komplexbildner zur Entfernung von Eisenverunreinigungen aus Industriemineralen mittels Bio-Flotation
- GaLlophore: Development of siderophore-based sorptive biocomposites for the recovery of Gallium
- Economic viability of gallium recovery and wastewater treatment.
- Technical challenges in removing iron from its complex with DFO.
- Importance of DFO cost in wastewater treatment.
- Difference in usability between DFO B and DFO E.
- DFO immobilisation and reusability of filters.
- Production scale considerations: If high DFO concentrations of 5 to 10 g/L can be achieved within a few days, separation between DFO-B and DFO-E variants is not as critical for wastewater treatment as it is for medicinal purposes, simplifying the purification process.
- Economic viability: The cost of siderophore production is the decisive factor determining feasibility and still present the primary barrier for wider industrial applications, including environmental engineering, agriculture, and metal recovery from waste streams.
- Immobilization: DFO immobilization offers a practical solution for industrial implementation, enabling efficient metal capture and allowing the material to be regenerated and reused multiple times, significantly reducing operational costs.
Main topics during the interview:
What did we learn?
Impact on our project: Dr. Jain's insights showed us that environmental applications where high yields and low costs matter more than pharmaceutical-grade purity, could also be a viable direction for our project. His emphasis on immobilization technology highlighted a practical pathway for industrial deployment that could make our Rhodococcus production system commercially viable for wastewater treatment and metal recovery applications.
Conclusion
Since the beginning of our project, expert consultations fundamentally reshaped our approach and broadened our understanding of DFO's potential applications. Prof. Julia Bandow provided crucial insights into analytical methods and regulatory considerations for our Rhodococcus expression system. Dr. Arno Cordes established that industrial viability requires yields of 10-100 g/L. Dr. Rohan Jain expanded our perspective beyond pharmaceutical applications, demonstrating that environmental applications such as wastewater treatment and metal recovery could benefit from DFO production, with the advantage that these applications are less constrained by purity requirements than pharmaceutical use.
These discussions revealed that while DFO's pharmaceutical applications for treating iron overload disorders remain valuable, the immediate path to industrial implementation may lie in environmental and metal recovery applications where cost and quantity, rather than pharmaceutical-grade purity, are the primary concerns. Our Rhodococcus-based production system addresses these priorities by offering a potentially cost-effective platform capable of utilizing diverse substrates and producing DFO at scale.
Recognising the limitations of our current work, we invite the iGEM and broader scientific audience to build upon this foundation, whether for developing cost-effective chelation therapies for patients with iron overload disorders, or creating sustainable solutions for metal recovery technologies from industrial waste streams.