Assessment of the Safety and Potential Allergenicity of a Subtilisin Enzyme Derived from Bacillus paralicheniformis AP-01 in Food Manufacturing Processes

The food enzyme subtilisin, classified under EC 3.4.21.62, is derived from the non-genetically modified Bacillus paralicheniformis strain AP-01, produced by Nagase (Europa) GmbH. It has been assessed to be free from viable production organism cells. The enzyme is intended for use in five distinct food manufacturing processes. As one of these processes effectively removes residual enzyme-Total Organic Solids (TOS), exposure calculations were conducted solely for the remaining four processes, resulting in an estimated maximum of 0.875 mg TOS/kg body weight per day among European populations.

The production strain is capable of producing bacitracin, which renders it non-compliant with the Qualified Presumption of Safety approach. While bacitracin was detected in the industrial fermentation medium, it was not present in the food enzyme. However, the detection limit for bacitracin was not adequate to definitively rule out the potential presence of bacitracin at a level that could pose a risk for antimicrobial resistance development.

A sequence comparison with known allergens revealed 28 matches with respiratory allergens, one with a contact allergen, and two with food allergens (specifically melon and pomegranate). The Panel acknowledged that the possibility of allergic reactions upon dietary exposure to this enzyme, particularly in individuals allergic to melon or pomegranate, could not be entirely ruled out, but it would not surpass the risks associated with consuming these fruits. Consequently, the safety of this food enzyme could not be conclusively established based on the available data.