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Geerlings et al. Microbiome Res Rep 2024;3:36 https://dx.doi.org/10.20517/mrr.2024.06 Page 13 of 15

synthetic media, high growth rates and biomass were observed.

In conclusion, we showed that pea peptone-based food-grade medium supplemented with glucose and
GlcNAc instead of mucin results in high biomass formation of A. muciniphila, which may be used for its
production for therapeutic purposes. The differences between growth on mucin and growth on glucose and
GlcNAc were shown at transcriptome and proteome levels. However, the general level 1 sKEGG
metabolism pathways showed high similarity between conditions and no major significant differences could
be identified. Furthermore, if minimal EPS production is required, the transcriptome data indicate that this
may be achieved by decreasing the GlcNAc concentration. Lastly, the use of synthetic medium affects the
cell morphology of A. muciniphila, resulting in elongated cells. Overall, our data suggest that the food-grade
synthetic medium composition described here could be used to produce A. muciniphila in high yields for
therapeutic purposes. This has recently been shown in the first proof-of-concept study with A.muciniphila
Muc grown on such a synthetic medium that showed the capacity of live and pasteurized cells to improve
T
[24]
several metabolic parameters in overweight and obese volunteers .

DECLARATIONS
Acknowledgments
We are grateful to Dr Bart Smit (NIZO) for a gift of industrial peptones and Dr Martin Pabst for giving
input in proteomics analysis. The content of the manuscript is derived from the dissertation.

Authors’ contributions
Designed experiments, conducted wet lab activities, analyzed data, constructed figures, and wrote the
manuscript: Geerlings SY
Conducted initial experiments and wrote the manuscript: van der Ark K
Processed raw data for transcriptomics and proteomics: Nijsse B, Boeren S
Provided input in result interpretation, guided the writing of the manuscript, as well as provided input and
critically reviewed the manuscript: van Loosdrecht M, Belzer C, de Vos WM

Availability of data and materials
Data supporting our findings are published as Supplementary Materials in this journal.

Financial support and sponsorship
This research was supported by the Netherlands Organization for Scientific Research (SIAM Gravity Grant
024.002.002).

Conflicts of interest
de Vos WM and Belzer C are inventors on patents related to the growth of Akkermansia spp. de Vos WM is
a co-founder and shareholder of the Akkermansia Company, commercializing pasteurized Akkermansia
muciniphila, while the other authors have declared that they have no conflicts of interest.

Ethical approval and consent to participate
Not applicable.

Consent for publication
Not applicable.