Abstract
Wheat is one of the most consumed foods in the world and unfortunately causes allergic reactions which have important health effects. The α-amylase/trypsin inhibitors (ATIs) have been identified as potentially allergen components of wheat. Due to a lack of data on optimization of ATI extraction, a new wheat ATIs extraction approach combining solvent extraction and selective precipitation is proposed in this work. Two types of wheat cultivars (Triticum aestivum L.), Julius and Ponticus were used and parameters such as solvent type, extraction time, temperature, stirring speed, salt type, salt concentration, buffer pH and centrifugation speed were analyzed using the Plackett-Burman design. Salt concentration, extraction time and pH appeared to have significant effects on the recovery of ATIs (p < 0.01). In both wheat cultivars, Julius and Ponticus, ammonium sulfate substantially reduced protein concentration and inhibition of amylase activity (IAA) compared to sodium chloride. The optimal conditions with desirability levels of 0.94 and 0.91 according to the Doehlert design were: salt concentrations of 1.67 and 1.22 M, extraction times of 53 and 118 min, and pHs of 7.1 and 7.9 for Julius and Ponticus, respectively. The corresponding responses were: protein concentrations of 0.31 and 0.35 mg and IAAs of 91.6 and 83.3%. Electrophoresis and MALDI-TOF/MS analysis showed that the extracted ATIs masses were between 10 and 20 kDa. Based on the initial LC-MS/MS analysis, up to 10 individual ATIs were identified in the extracted proteins under the optimal conditions. The positive implication of the present study lies in the quick assessment of their content in different varieties especially while considering their allergenic potential.
Highlights
Wheat has become one of the most important foods in the world
About 0.16 ± 0.03 to 0.61 ± 0.04 mg/mL of the protein concentration, and 26.6 ± 1.6 to 90.3 ± 2.7% of inhibition of amylase activity (IAA) were registered with Ponticus
It appears that the type of solvent mixture (X1), the composition of the solvent (X2) as well as the ratio of sample/solvent (X3) had no significant effect on the protein content and IAA as noted for Julius and Ponticus. These results show that dichloromethane can be used with the same performance as chloroform for carrying out amylase/trypsin inhibitors (ATIs) extraction
Summary
Wheat has become one of the most important foods in the world. Its growing consumption has led to an increase in allergies such as gluten intolerance, celiac disease and IgE-mediated allergies (including respiratory immune responses, especially for bakers) that affects about 0.4–2% of the general population, depending on the age and region [1,2,3]. A food allergy is described as an adverse health effect involving immunological mechanisms (allergic reactions) that can be induced in sensitized children and adults following dietary exposure to relevant allergens in foods [4] These allergens are usually proteins, but sometimes chemicals such as haptens have been reported [5]. Newer studies have explored other wheat compounds that may cause such problems and one of these candidates are α-amylase/trypsin inhibitors (ATIs)
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