The intended users

By engaging with various stakeholders as part of our human practices work, we identified a clear niche for our solution in the biotechnology industry. Worldwide firms such as COFCO (China), DSM (The Netherlands), Genencor, A Danisco Division (USA), Novozymes A/S (Denmark), RZBC Group, (China), Zymergen (USA), etc. (Cairns et al. (2018)) commonly use A. niger as a production organism for various organic acids and enzymes but they require specialised growth media.

These companies could likely be interested in developing new A. niger production strains based on our furfural detecting and converting strain (Martins-Santana L, et al.(2018), Walker et al. (2018), Amer et al. (2021)). This would open up for cheaper production capabilities, as the growth media would be based on lignocellulosic waste instead of defined media (Gwehenberger et al. (2008), Woodley et al. (2013), Sheldon et al. (2018)). Simultaneously, it would aid in switching to a more sustainable production for these companies.

Our challenges

The project is still at an early stage, implementation-wise. Currently, we are working on a proof of concept level, meaning we still need to work on up-scaling before we have a strain that we can supply to corporations, as exemplified in figure 1. Up-scaling experiments would include testing our strain on lignocellulosic waste. If successful, we would start investigating fermentations with increasing volumes, and optimising our strain, possibly through continuous fine-tuning of the promoter system and furfural converting protein.

Challenges in large-scale bioproduction rapidly grow with cost of equipment, personnel, and time required. Therefore, a cost-benefit analysis would be required to assess the efficiency of the production strain for the industry (Al-onso et al. (2017)).

Figure 1. An overview of the development and Implementation of our A. niger strain. We have completed several rounds of the Design-Build-Test-Learn (DBTL) during the ideation and proof of concept steps of our project. The next step is to test the scalability of the project including small-scale fermentation experiments and pilot-scale fermentation. These will pave the way for upscaling to industrial applications.

The issue of safety

Many of the safety concerns that apply to our production strain are the same that apply to working with WT A. niger. The organism is categorised as Generally Regarded As Safe (GRAS), which means that it is a safe organism to work with, as long as the correct safety standards are upheld. The laboratory experiments have thus been performed to fulfil the Good Laboratory Practices and biosafety rules as described in our safety tab. To confirm that our final strain is as safe to work with as WT A. niger, screening is needed to confirm that our furfural detecting strain does not have an increased pathogenicity (something we definitely do not expect to be the case).

Summary

We envision our production strain bringing great value to firms that frequently use A. niger as a production organism for their pipelines. Our A. niger strain is designed to convert renewable feedstock into chemicals, contributing to the transition from an economy based on fossil fuels to a circular bio-based one. Further work and research need to be done to achieve a strain that can be widely used on an industrial-level scale, however we believe that this is an attainable goal.

References

• Al-onso, D. M., et al. Increasing the revenue from lignocellulosic biomass: Maximizing feedstock utilization. Science advances, 3(5) (2017)
• Amer, B. & Baidoo, E. Omics-Driven Biotechnology for Industrial Applications. Frontiers in Bioengineering and biotechnology 9, 613307 (2021)
• Cairns, T. C., et al. How a fungus shapes biotechnology: 100 years of Aspergillus niger research. Fungal biology and biotechnology, 5, 13 (2018)
• Gwehenberger, G. & Narodoslawsky, M. Sustainable processes—The challenge of the 21st century for chemical engineering. Process Safety and Environmental Protection 86. p. 321-327 (2008)
• Martins-Santana L, et al. Systems and synthetic biology approaches to engineer fungi for fine chemical production. Frontiers in bioengineering and biotechnology, 6, 117 (2018)
• Sheldon, R. A., & Woodley, J. M. Role of Biocatalysis in Sustainable Chemistry. Chemical reviews, 118(2), 801–838 (2018)
• Walker, R. S. K. & Pretorius, I. S. Applications of yeast synthetic biology geared towards the production of biopharmaceuticals. Genes, 9(7): 340 (2018)
• Woodley, J. et al. A future perspective on the role of industrial biotechnology for chemicals production. Chemical Engineering Research and Design 91. p. 2029-2036. (2013)