Safety evaluation of honey truffle sweet protein produced from Komagataellaphaffii

Honey truffles (Mattirolomyces terfezioides) contain a natural protein which exhibits intense sweetness. This protein is produced via precision fermentation using Komagataella phaffii (formerly Pichia pastoris) as the chassis and is processed into a spray dried powder referred to as Honey Truffle Sweet Protein (HTSP) and the powder contains at least 50% w/w total protein, of which at least 50% is the active sweet protein, Honey Truffle Active Component (HT-AC). HT-AC is 500-2500 times sweeter than sucrose dependent on food/beverage formulations. A set of genotoxicity assays and oral repeated dose studies were conducted to assess the safety of HTSP for its use as a sweetener in food and beverages. The results of the in vitro bacterial mutagenicity and mammalian cell micronucleus tests demonstrated a lack of genotoxic effects. In the 13-week oral exposure study, male and female Wistar Han rats were dosed by gavage to 1800, 3600, and 5400 mg/kg/day of HTSP (equivalent to approximately 500, 1000 and 1500 mg/kg/day of the HT-AC). Treatment was well tolerated with neither mortality nor HTSP-related adverse clinical pathology findings, organ weight effects, or macroscopic or microscopic observations. Consequently, the no observed adverse effect level (NOAEL) was 5400 mg/kg/day (1500 mg/kg/day HT-AC). The results supports the safety of HTSP for use as a dietary sweetener in food and beverage applications.