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The world's hunger for novel food ingredients drives the development of safe, sustainable, and nutritious novel food products. For foods containing novel proteins, potential allergenicity of the proteins is a key safety consideration. One such product is a fungal biomass obtained from the fermentation of Rhizomucor pusillus. The annotated whole genome sequence of this strain was subjected to sequence homology searches against the AllergenOnline database (sliding 80-amino acid windows and full sequence searches). In a stepwise manner, proteins were designated as potentially allergenic and were further compared to proteins from commonly consumed foods and from humans. From the sliding 80-mer searches, 356 proteins met the conservative >35% Codex Alimentarius threshold, 72 of which shared ≥50% identity over the full sequence. Although matches were identified between R. pusillus proteins and proteins from allergenic food sources, the matches were limited to minor allergens from these sources, and they shared a greater degree of sequence homology with those from commonly consumed foods and human proteins. Based on the in silico analysis and a literature review for the source organism, the risk of allergenic cross-reactivity of R. pusillus is low., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Kevin Scaife reports financial support was provided by The Protein Brewery B.V. Lucie Pařenicová reports financial support was provided by The Protein Brewery B.V. Trung Vo reports financial support was provided by The Protein Brewery B.V. Elisa Leune reports financial support was provided by The Protein Brewery B.V. Steve Taylor reports financial support was provided by The Protein Brewery B.V. Richard Goodman reports financial support was provided by The Protein Brewery B.V. Mohamed Abdelmoteleb declares that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Yvonne Dommels reports financial support was provided by The Protein Brewery B.V. Kevin Scaife reports a relationship with The Protein Brewery B.V. that includes: consulting or advisory. Lucie Parenicova reports a relationship with The Protein Brewery B.V. that includes: employment. Trung Vo reports a relationship with The Protein Brewery B.V. that includes: consulting or advisory. Elisa Leune reports a relationship with The Protein Brewery B.V. that includes: employment. Steve Taylor reports a relationship with The Protein Brewery B.V. that includes: consulting or advisory. Richard Goodman reports a relationship with The Protein Brewery B.V. that includes: consulting or advisory. Yvonne Dommels reports a relationship with The Protein Brewery B.V. that includes: employment., (Copyright © 2024 Elsevier Inc. All rights reserved.)