Analyzing pepsin degradation assay conditions used for allergenicity assessments to ensure that pepsin susceptible and pepsin resistant dietary proteins are distinguishable

• DOI: 10.1371/journal.pone.0171926
• Publication Date: 2017-02-16T21:15:21Z
• Authors: Rong Wang; Thomas C. Edrington; S. Bradley Storrs; Kathleen S. Crowley; Jason M. Ward; Thomas C. Lee; Zi L. Liu; Bin Li; Kevin C. Glenn
• Keywords: Pepsin; Proteolysis

The susceptibility of a dietary protein to proteolytic degradation by digestive enzymes, such as gastric pepsin, provides information on the likelihood systemic exposure structurally intact and biologically active macromolecule, thus informing safety proteins for human animal consumption. Therefore, purpose standardized in vitro studies that are performed during assessments is distinguish whether interest susceptible or resistant pepsin via study design enables study-to-study comparison. Attempting assess under wide-range possible physiological conditions poses problem because lack robust consistent data collected large-range sub-optimal conditions, which undermines needs harmonize conditions. This report systematically compares effects pH, incubation time, pepsin-to-substrate ratio relative five proteins: three [ribulose 1,5-bisphosphate carboxylase-oxygenase (Rubisco), horseradish peroxidase (HRP), hemoglobin (Hb)], two [lipid transfer (LTP) soybean trypsin inhibitor (STI)]. results indicate readily distinguishable from pepsin-resistant when reaction within well-characterized optima pepsin. current assay with low pH high fits this purpose. Using non-optimal and/or ratios resulted no longer being reliably degraded stomach enzyme, compromises ability between and, therefore, providing useful an overall weight-of-evidence approach assessing proteins.