A paper titled "Determination of bacillopeptidase F, a protein produced by Bucillus subtilis natto using ELISA" has been published

A paper titled "Determination of bacillopeptidase F, a protein produced by Bucillus subtilis natto using ELISA" has been published

Daiwa Pharmaceutical developed an enzyme-linked immunosorbent assay (ELISA) to determine bacillopeptidase F in Daiwa's functional food material “NKCP” and published the details in the Journal of Nutritional Food, Vol. 9, No. 3/4, (2006) (released by Japan Health Food & Nutrition Food Association.) Daiwa Pharmaceutical and Dokkyo Medical University jointly performed this study with the support of research institutes including the Japan Food Research Laboratory.

Functional ingredients in food should be defined for safety and efficacy evaluations. Nowadays, it is regarded important to perform quantitation in addition to qualitative assay.

Bacillopeptidase F is a serine protease produced by Bacillus subtilis natto, which has a different amino acid sequence to subtilisin, conventionally called nattokinase. In previous studies, “NKCP” containing bacillopeptidase F was shown to have (1) fibrinolytic, (2) anticoagulant and (3) blood viscosity reducing effects (triple functions), and these ingredients need to be discriminated and determined correctly and easily.

The widely used quantitative assay for Bucillus subtilis natto cultures determines fibrin degraded by protease, using fibrin as the substrate. However, as various proteases degrade fibrin, it is impossible to differentiate proteases derived from Bacillus subtilis natto from other proteases when they are mixed. In contrast, the new ELISA method, developed by Daiwa Pharmaceutical, allows determination of bacillopeptidase F only. Validation of an ELISA kit for quantitation was performed in cooperation with the Japan Food Research Laboratory. The results suggest that the kit is available for determination of the functional ingredient bacillopeptidase F.

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