In vitro digestibility and immunoreactivity of thermally processed peanut

Figures & data

Figure 1. SDS-PAGE analysis of simulated oral (A), oral-gastric (B,C) and oral-gastric-intestinal (D,E) digestion of raw peanut. R – undigested raw peanut extract; C – chew sample; G – no enzyme gastric control; I – no enzyme intestinal control; Panels B and C Lanes 1–6 are gastric digestion time points – 0.3, 2, 10, 40, 70 and 120 min; Panels D and E Lanes 1–4 are intestinal digestion time points – 5, 15, 30 and 120 min.

Table 1. Proteins identified in gels by tryptic digestion (bands annotated in Figures 1(B ) and 3(B)).

Band	Accession	Description	Coverage	Peptides	Unique peptides	MW [kDa]	Area
1	Q0GM57	Ara h 3	33.0	13	11	58.2	5.61E + 09
1	Q647H4	Ara h 3	30.6	14	5	61.5	2.99E + 09
2	Q6PSU2	Ara h 2	41v3	11	11	20.1	1.23E + 09

Figure 2. SDS-PAGE analysis of simulated oral (A), oral-gastric (B,C) and oral-gastric-intestinal (D,E) digestion of roasted peanut. R – undigested roasted peanut extract; C – chew sample; G – no enzyme gastric control; I – no enzyme intestinal control; Panels B and C Lanes 1–6 are gastric digestion time points – 0.3, 2, 10, 40, 70 and 120 min; Panels D and E Lanes 1–4 are intestinal digestion time points – 5, 15, 30 and 120 min.

Figure 3. SDS-PAGE analysis of simulated oral (A), oral-gastric (B,C) and oral-gastric-intestinal (D,E) digestion of boiled peanut. R – undigested boiled peanut extract; C – chew sample; G – no enzyme gastric control; I – no enzyme intestinal control; Panels B and C Lanes 1–6 are gastric digestion time points – 0.3, 2, 10, 40, 70 and 120 min; Panels D and E Lanes 1–4 are intestinal digestion time points – 5, 15, 30 and 120 min.

Figure 4. Immunoblotting analysis of the soluble fraction of digested peanut and processing peanut. Immunoblots were developed using polyclonal antibody preparations to Ara h1, Ara h3 and Ara h 2/6. The order of the samples in lane 1: non-digested raw peanut; lanes 2, 4 and 6: soluble fractions after 10 min gastric digestion of raw peanut, roasted peanut and boiled peanut, respectively; lanes 3, 5 and 7: soluble fractions after 120 min gastric digestion of raw peanut, roasted peanut and boiled peanut, respectively.

Figure 5. IgE immunoreactivity of digested peanut and processing peanut using plasma from peanut-allergic patients. The order of the samples in lane 1: non-digested raw peanut; lanes 2, 4 and 6: soluble fractions after 10 min gastric digestion of raw peanut, roasted peanut and boiled peanut, respectively; lanes 3, 5 and 7: soluble fractions after 120 min gastric digestion of raw peanut, roasted peanut and boiled peanut, respectively.

Figure 6. Comparison of the IgE-binding capacity of oral-gastric digesta. (a) samples were digested for 10 min, (b) samples were digested for 120 min and (c) IC50 values of all digesta.