Safety Evaluation of Fermotein: Allergenicity, Mycotoxin Production, Biochemical Analyses and Microbiology of a Fungal Single-cell Protein Product

Aim: Single-cell proteins (SCPs) are considered as innovative and sustainable alternatives to animal-based products. Fermotein is an innovative SCP obtained from fermentation of the filamentous fungus Rhizomucor pusillus. The toxicity, capability to produce secondary metabolites and allergenic potential of this fungus has never been assessed before. Like other filamentous fungi, there is a lack of information on this species to assess its safety for human consumption. The objective of the current study was to investigate the safety of Fermotein and its source Rhizomucor pusillus regarding toxicity, capability to produce secondary metabolites and allergenicity. In addition, possible contaminants were also examined. Methodology: The genome of Rhizomucor pusillus was sequenced and annotated in order to screen for production of common mycotoxins, antibiotic synthesis pathways, mucormycosis-related virulence factors and in silico potential cross-reactivity with known food allergens. The presence of mycotoxins and allergens were validated by laboratory analysis. The level of RNA, heavy metals and microbiological contaminants were also determined. Results: No mycotoxin production-related genes were identified in the genome of Rhizomucor pusillus nor were mycotoxins found in Fermotein. Six proteins present in Fermotein showed high homology with five known food allergens. No gene clusters were found that corresponded with antibiotic synthesis pathways. Although 10 proteins in the genome of Rhizomucor pusillus may represent mucormycosis-related virulence factors, no cases of mucormycosis after oral intake are reported. The level of heavy metals and microbiological contaminants were below legislative limits, whereas RNA content was 4.9 ± 0.2% of dry matter. Conclusion: No safety concerns were identified for Fermotein or its source Rhizomucor pusillus, except the potential for cross-reactivity with five known food allergens. This should be taken into account for communication with consumers. Information from the current study contributes to the body of evidence for determination of Qualified Presumption of Safety status of Rhizomucor pusillus.