Clinostat Rotation Affects Metabolite Transportation and Increases Organic Acid Production by Aspergillus carbonarius , as Revealed by Differential Metabolomic Analysis.

Contamination by fungi may pose a threat to the long-term operation of the International Space Station because fungi produce organic acids that corrode equipment and mycotoxins that harm human health. Microgravity is an unavoidable and special condition in the space station. However, the influence of microgravity on fungal metabolism has not been well studied. Clinostat rotation is widely used to simulate the microgravity condition in studies carried out on Earth. Here, we used metabolomics differential analysis to study the influence of clinostat rotation on the accumulation of organic acids and related biosynthetic pathways in ochratoxin A (OTA)-producing Aspergillus carbonarius. As a result, clinostat rotation did not affect fungal cell growth or colony appearance but significantly increased the accumulation of organic acids, particularly isocitric acid, citric acid, and oxalic acid, and OTA both inside cells and in the medium, as well as resulted in a much higher level of accumulation of some products inside than outside cells, indicating that the transport of these metabolites from the cell to the medium was inhibited. This finding corresponded to the change in the fatty acid composition of cell membranes and the reduced thickness of the cell walls and cell membranes. Amino acid and energy metabolic pathways, particularly the tricarboxylic acid cycle, were influenced the most during clinostat rotation compared to the effects of normal gravity on these pathways. IMPORTANCE Fungi are ubiquitous in nature and have the ability to corrode various materials by producing metabolites. Research on how the space station environment, especially microgravity, affects fungal metabolism is helpful to understand the role of fungi in the space station. This work provides insights into the mechanisms involved in the metabolism of the corrosive fungus Aspergillus carbonarius under simulated microgravity conditions. Our findings have significance not only for preventing material corrosion but also for ensuring food safety, especially in the space environment.