Proteolysis by Lactobacillus fermentum IFO3956 isolated from Egyptian milk products decreases immuno-reactivity of alpha(S1)-casein

Proteinase activity of Lactobacillus fermentum IFO3956 cells was higher when they were grown on milk-based media than on 10\% reconstituted skim milk. The lowest protease activity was observed when cells were grown on milk-free media. The extraction of milk-induced cell-bound proteases from Lb. fermentum IFO3956 was most efficient using 1\% Tween 80 while the use of 1\% SDS inhibited all proteolytic activity. Two bands of similar to 35 and > 100 kDa were observed by zymogram, indicating that proteolytic activity corresponded to the presence of at least two types of enzymes or two molecular forms of one enzyme. Mass spectrometry analyses of alpha(S1)-casein hydrolysates detected 24 peptides with sizes ranging from 5 to 36 amino acids, including 9 phosphorylated peptides, resulting from the fermentation of Lb. fermentum IFO3956 of aS1-casein. Most of the identified peptides originated from the N-terminal portion of alpha(S1)-casein. The studied bacterial strain could hydrolyze alpha(S1)-casein in many sites including the epitopes triggering the allergic reactions against alpha(S1)-casein e. g. at the positions 23, 30, 41, 71, 91, 98, 126, 179. After hydrolysis of alpha(S1)-casein with Lb. fermentum IFO3956 the recognition and the binding of this casein to IgE from the pooled sera of 18 patients with cow's milk allergy was significantly reduced.