Safety evaluation of food enzymes produced by a safe strain lineage of Bacillussubtilis

The safety of microbially-derived food enzymes must be carefully assessed before market introduction. The production strain's safety is central to the assessment. In this paper, we have determined that DSM's Bacillus subtilis strain lineage can be considered safe for food enzyme production. The mutations introduced into this non-pathogenic and non-toxigenic microorganism do not lead to any safety concerns, as ensured by a thorough characterization of the strain lineage. The safety of both targeted and randomly introduced changes into the production strain's genome is confirmed by validating the absence of vector sequences and antibiotic resistance genes in all relevant production strains, and by demonstrating absence of cytotoxic peptide production. Furthermore, three food enzyme preparations produced by strains within this lineage did not show genotoxic potential. 90-day oral toxicity studies performed with the same enzyme preparations did not reveal toxicologically significant adverse effects. These results demonstrate absence of safety concerns from the introduced genetic modifications. Based on the establishment of this safe strain lineage, we postulate that future enzymes produced by current and new strains derived from the lineage can be safely developed without additional genotoxicity and systemic toxicity studies, allowing for a reduction of animal testing without compromising on product safety. •Bacillus subtilis is recognized as a safe microorganism for enzyme production.•Random mutations applied into this safe strain lineage do not lead to a safety risk.•Genetic engineering within this safe strain lineage did not lead to safety risks.•Safety of DSM's Bacillus subtilis food enzymes was confirmed with toxicity studies.•Safety of new enzymes from this lineage can be established without new safety studies.