ELISA analysis of soybean trypsin inhibitors in processed foods.
Soybean proteins are widely used in human foods in a variety of forms, including infant formulas, flour, protein concentrates, protein isolates, soy sauces, textured soy fibers, and tofu. The presence of inhibitors of digestive enzymes in soy proteins impairs the nutritional quality and possibly the safety of soybeans and other legumes. Processing, based on the use of heat or fractionation of protein isolates, does not completely inactivate or remove these inhibitors, so that residual amounts of inhibitors are consumed by animals and humans. New monoclonal antibody-based immunoassays can measure low levels of the soybean Kunitz trypsin inhibitor (KTI) and the Bowman-Birk trypsin and chymotrypsin inhibitor (BBI) and the Bowman-Birk foods. The enzyme-linked immunosorbent assay (ELISA) was used to measure the inhibitor content of soy concentrates, isolates, and flours, both heated and unheated; a commercial soy infant formula; KTI and BBI with rearranged disulfide bonds; browning products derived from heat-treatment of KTI with glucose and starch; and KTI exposed to high pH. The results indicate that even low inhibitor isolates contain significant amounts of specific inhibitors. Thus, infants on soy formula consume about 10 mg of KTI plus BBI per day. The immunoassays complement the established enzymatic assays of trypsin and chymotrypsin inhibitors, and have advantages in (a) measuring low levels of inhibitors in processed foods; and (b) differentiating between the Kunitz and Bowman-Birk inhibitors. The significance of our findings for food safety are discussed.
Brandon
D L
DL
Food Safety Research Unit, USDA Agricultural Research Service, Albany, CA 94710.
Bates
A H
AH
Friedman
M
M
D000468:Alkalies / Q000494:pharmacology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D002241:Carbohydrates / Q000494:pharmacology; D010450:Endopeptidases / Q000032:analysis / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D005504:Food Analysis; D005511:Food Handling; D006358:Hot Temperature; D006801:Humans; D007223:Infant; D007225:Infant Food / Q000032:analysis; D014674:Plant Proteins, Dietary / Q000032:analysis*; D030262:Soybean Proteins; D013025:Glycine max / Q000201:enzymology*; D014358:Trypsin Inhibitor, Bowman-Birk Soybean / Q000032:analysis / Q000276:immunology; D014360:Trypsin Inhibitor, Kunitz Soybean / Q000032:analysis / Q000187:drug effects / Q000276:immunology; D014361:Trypsin Inhibitors / Q000032:analysis* / Q000276:immunology
D000468:Alkalies; D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D000939:Epitopes; D014674:Plant Proteins, Dietary; D030262:Soybean Proteins; D014358:Trypsin Inhibitor, Bowman-Birk Soybean; D014361:Trypsin Inhibitors; D014360:Trypsin Inhibitor, Kunitz Soybean; D010450:Endopeptidases