Comparative study of protease hydrolysis reaction demonstrating Normalized Peptide Bond Cleavage Frequency and Protease Substrate Broadness Index

    Abstract: Two commercial proteases (subtilisin-typed FNA from Bacillus amyloliquefaciens, and chy-motrypsin-like NPP from Nocardiopsis prasina), porcine pepsin, porcine pancreatin having protease activity and their combinations were studied in vitro by LC-MS for their ability to digest soy protein isolate (SPI) under conditions close to those found in the stomach (pH 3.7) and small intestine (pH 6.5). The total number of peptides generated, and their size dis-tribution were obtained under each set of the digestion conditions. These peptides were grouped according to their C-terminal amino acid (AA) residue (P1) and mass, based on which two concepts were proposed, i.e., Normalized Peptide Bond Cleavage Frequency (NPBCF) and Protease Substrate Broadness Index (PSBI). At pH 3.7, FNA+pepsin increased PSBI vs. pepsin alone by 2.7 and 4.9 percentage points (p.p.) at a SPI:protease ratio of 20:1 and 100:1, respectively. At pH 6.5, FNA+pancreatin improved PSBI by 9.1 and 10.2 p.p. at SPI:protease 20:1 and 100:1, respectively, vs. pancreatin alone. NPP generated 38% more peptides than FNA when administered with pancreatin at SPI:protease 200:1:1 and pH 6.5, but FNA alone (28.9) or FNA+pancreatin (29.1) gave a higher PSBI than pan-creatin (22.2), NPP (20.3) and NPP+pancreatin (22.0). At pH 3.7 FNA generated 59% and 39% of peptides of pepsin at SPI:protease of 20:1 and 100:1, respectively, and both groups of peptides had similar size distribution. At pH 6.5 more small sized peptides were generated by FNA or FNA+pancreatin than pancreatin and NPP alone or pancreatin+NPP. In conclu-sion, FNA showed complementary effects with pepsin and pancreatin in terms of PSBI and generated more small sized peptides compared to NPP.
    Document information
    Product / service: Carbohydrases and Proteases
    Publication date: 22/09/2020
    Species: Multi-species
    Authors: Shukun Yu, PhD
    Doctype: Publications & Citations
    Publication / conference: Plos One
    Regions and countries:
    Keywords: protease, protein
    Production challenge(s):
    Diets: All diets
    Brands: Axtra® PRO
    Resource ref:
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