In vitro digestibility of bovine β-casein with simulated and human oral and gastrointestinal fluids. Identification and IgE-reactivity of the resultant peptides.
Stability during digestion is considered an important feature in determining the allergenicity of food proteins. This study aimed to provide an immunological characterisation of the digestion products of the major cow's milk allergen β-casein (β-CN) produced by in vitro orogastrointestinal hydrolysis with simulated and human digestive fluids. β-CN was unaffected by oral digestion, but quickly broke down during the early stages of gastric digestion. The degradation with human fluids was faster than that with commercial enzymes. There were similarities in the peptide patterns of the hydrolysates produced in both models, showing 20 peptides in common after gastric digestion. After gastroduodenal digestion, the human fluids gave less numerous and shorter peptides. The IgE binding of most of the individual sera used to the hydrolysates produced with simulated and human fluids increased at the end of the gastric phase and decreased when the duodenal digestion was completed. Two IgE-binding synthetic peptides: β-CN (57-68) and β-CN (82-93), which matched fragments released by β-CN following in vitro digestion with simulated and human fluids, consisted of the most immunoreactive areas of the protein. The similarities found between the in vitro simulated digestion system and that using human digestive fluids suggest that the former would provide a reasonably good estimation of the potential allergenicity of protein digests.
Instituto de Investigación en Ciencias de la Alimentación (CIAL), CSIC-UAM, Madrid, Spain.
Sara
S
Benedé
Iván
I
López-Expósito
Gustavo
G
Giménez
Galina
G
Grishina
Ludmilla
L
Bardina
Hugh A
HA
Sampson
Elena
E
Molina
Rosina
R
López-Fandiño
D000485:Allergens / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D000818:Animals; D002364:Caseins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D002417:Cattle; D004063:Digestion; D005512:Food Hypersensitivity / Q000276:immunology / Q000378:metabolism*; D005750:Gastric Juice / Q000378:metabolism*; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology*; D008892:Milk / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D008954:Models, Biological; D010455:Peptides / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D012463:Saliva / Q000378:metabolism*
D000485:Allergens; D002364:Caseins; D010455:Peptides; D007073:Immunoglobulin E
Human digestive fluids; IgE; In vitro digestion; Milk allergy; Peptide microarray; β-Casein