The European Food Safety Authority (EFSA) concluded that the food enzyme thermolysin, produced with the non-genetically modified bacterium Anoxybacillus caldiproteolyticus strain DP-Fzj32 by Danisco US Inc., does not raise safety concerns for infants, toddlers, adolescents, adults and the elderly under the intended conditions of use. However, the margin of exposure was insufficient to exclude safety concerns for children, according to a scientific opinion adopted on 20 May 2026 and published on 11 June 2026.
The opinion assessed thermolysin (EC 3.4.24.27), a protease that hydrolyses peptide bonds in proteins, for use in three food manufacturing processes: production of modified milk proteins, production of protein hydrolysates from meat and fish, and production of protein hydrolysates from plants and fungi. Dietary exposure was estimated at up to 0.874 mg total organic solids (TOS) per kg body weight per day across European populations. Genotoxicity tests showed no safety concern, and a 90-day oral toxicity study in rats identified a no observed adverse effect level (NOAEL) of 108 mg TOS/kg bw per day, the highest dose tested. Margins of exposure ranged from 124 for children to 2348 for the elderly, but the panel considered the margin for children (124) insufficient to exclude safety concerns. A search for amino acid sequence homology to known allergens found no match, and the panel assessed the likelihood of allergic reactions as low.
This is the first EFSA safety evaluation of thermolysin from this specific strain, submitted under Regulation (EC) No 1332/2008 on food enzymes. The application was part of a batch of five food enzyme applications submitted by Danisco US Inc. and Amano Enzyme Inc., requested by the European Commission. The opinion follows EFSA's updated guidance documents, including the 2021 Scientific Guidance for the submission of dossiers on food enzymes and the 2023 guidance on food manufacturing processes for exposure assessment. The production strain was identified as a natural isolate, and no antimicrobial resistance genes of concern were found, though the panel noted a limitation: no whole genome sequence-based analysis of virulence or toxin genes was provided.
Stakeholder impact: For Danisco US Inc., the positive safety conclusion for most age groups supports the company's application for EU market authorisation, but the flagged concern for children may require additional data or restricted use conditions. For EU food manufacturers, the enzyme could be used to produce protein hydrolysates for dairy, meat, fish, and plant-based products, potentially improving texture or nutritional value, but the children safety gap may limit its use in products intended for that age group. For consumers, the opinion indicates no safety concern for the general population, but parents and caregivers of children may face uncertainty until the issue is resolved. For EU regulators, the opinion provides a basis for the Commission to decide on inclusion in the EU Community list of authorised food enzymes, possibly with specific conditions for children's products.
Expected follow-up: The European Commission will consider EFSA's opinion in the authorisation process. The applicant may be asked to provide additional data to address the children safety concern, or the authorisation may include restrictions on use in foods targeted at children.