Honey truffle sweetener (HTS), a
121 amino acid protein is identified
as a high-intensity sweetener found naturally occurring in the Hungarian
Sweet Truffle Mattirolomyces terfezioides, an edible mushroom used in regional diets. The protein is intensely
sweet, but the truffle is difficult to cultivate; therefore, the protein
was systematically characterized, and the gene coding for the protein
was expressed in a commonly used host yeast Komagataella
phaffii. The heterologously expressed protein maintained
the structural characteristics and sweet taste of the truffle. Preliminary
safety evaluations for use as a food ingredient were performed on
the protein including digestibility and in silico approaches for predicting the allergenicity and toxicity of the
protein. HTS is predicted to be nonallergenic, nontoxic, and readily
digestible. This protein is readily produced by precision fermentation
of the host yeast, making it a potential replacement for both added
sugars and small molecule high-intensity sweeteners in food.