Cleavage specificity of the pitcher fluid proteases from Nepenthes × miranda and their reduction on allergenicity of cow's milk proteins

In this study, the pitcher fluid proteases from Nepenthes × miranda were researched as a novel protease resource due to their cleavage specificity and ability to reduce the allergenicity of cow's milk proteins. We found that these proteases are particularly efficient at the P1 position with K, L, V, S, I, and R residues and exhibit similar preferences to amino acid residues at the P1′ position. It is concluded that P1 is responsible for specificity of pitcher fluid proteases, while P1′ tends to show their broadness when hydrolyzation happens. And consistent with the destroying of epitopes, in vivo assays also demonstrated a reduction in allergenicity from both whey protein concentrates and caseins, although the effect on caseins paled to whey protein concentrates. Therefore, these proteases hold significant potential and warrant further development for applications addressing cow's milk protein allergies.