Role of pepsin in modifying the allergenicity of bhetki (Lates calcarifer) and mackerel (Rastrelliger kanagurta) fish (original) (raw)
Related papers
Identification and Characterization of Main Allergic Proteins in Cooked Wolf Herring Fish
Iranian journal of allergy, asthma, and immunology, 2016
Our aim in this study was to identify and characterize allergic proteins in cooked wolf herring fish. We heated the crude extract alternatively at 50, 60, 70, 80, 90, and 100°C for one hour and results were compared by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Also, proteins were immunoblotted with fish-sensitive patients' sera. The major allergenic proteins were identified via mass spectrometry. These allergenic proteins were then purified by anion exchange chromatography and the IgE-immunoreactivity of the fractions was compared with the crude extracts via disk enzyme-linked immunosorbent assay (ELISA). SDS-PAGE of the crude extract showed more than 15 distinct protein bands. Five of these proteins, with apparent molecular weights of 12, 18, 24, 38, and 51 kDa, were only observed in the 100°C heated extract. Immunoblotting of the heated extract revealed that the 12 and 51 kDa proteins were IgE-immunoreactive with 88 percent of fish-sensitive patient...
Journal of the …, 2011
Food allergy is mediated by IgE antibodies to food components, usually proteins, and the digestion resistance of each protein is considered to be one of important parameters for its allergenic potential. 1-4) The safety of foods derived from genetically modified plants must be carefully assessed, especially in regard to food allergy. 5-12) The International Food Biotechnology Council and the International Life Sciences Institute jointly developed a step-wise approach to assess the safety of newly expressed proteins. 12) Their recommendations are based on similarities to known protein allergens and resistance to digestion of the proteins. However, the digestibility of a protein measured by the in vitro assay in simulated gastric fluid (SGF) or simulated intestinal fluid (SIF) is greatly influenced by the assay conditions. 13-15) The ratio of pepsin (or pancreatin) to food proteins during the digestion process significantly affects the rate of digestion. Thus, evaluation of relative digestion resistance of various allergenic and non-allergenic proteins under standardized conditions is very useful to obtain the relationship between allergenic potential and digestibility. In addition, the allergenic potential of food is altered by various manufacturing processes, 16) and also the digestibility of food proteins can be affected by food processing, for example, heating, chemical-treatment, exposure to high-pressure, fermentation, etc. Since little is known about changes in the digestibility as a result of food processing, we also investigated the reduction in digestion resistance by preheating, the most popular processing method. We tested five food allergens, ovalbumin (OVA), ovomucoid (OVM), b-lactoglobulin (BLG), bovine serum albumin (BSA), soybean trypsin inhibitor (STI). We also studied the digestibility of four food proteins, horseradish peroxidase (HRP), ribulose-1,5-bisphosphate carboxylase/oxidase (RBC), phosphinothricin acetyltransferase (PAT) and corn storage protein, zein, whose allergenicity have not been reported. A plant lectin, concanavalin A (Con A) was investigated for reference. MATERIALS AND METHODS Materials Pepsin (catalog number P6887), pancreatin (catalog number P8096), and the test proteins were purchased from Sigma Chemical Co. (St. Louis, MO, U.S.A.) in the purest form available. The concentrations of all test proteins except PAT, RBC and zein were 5 mg/ml water for SGF and 2 mg/ml water for SIF. PAT and RBC were dissolved in 50 mM Tris-HCl (pH 9.5) and zein was dissolved in 10% dimethylsulfoxide (final concentration in the test solution was 0.5%). Gels and reagents for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) were purchased from Invitrogen (Carlsbad, CA, U.S.A.). Preparation of SGF and SIF Pepsin (3.8 mg; approximately 13148 units of activity) was dissolved into the 5 ml gastric control solution (G-con; 2 mg/ml NaCl, pH adjusted to 2.0 with dilute HCl) and the activity of each newly prepared SGF solution was defined as production of a DA280 of 0.001 per min at pH 2.0 at 37°C measured as trichloro-acetic acid (TCA)-soluble products with hemoglobin as substrate. SIF was prepared as described in the United States Pharmacopoeia, 24th edition, p2236, and 10 mg/ml pancreatin was dissolved in intestinal control solution (I-con; 0.05 M KH 2 SO 4 , pH 6.8). Both solutions were used within the same day. Incubation in SGF or SIF SGF (1520 ml) was incubated at 37°C for 2 min before addition of 80 ml of test protein solution (5 mg/ml) at time zero. At each scheduled time point (0.5, 2, 5, 10, 20, 30, 60 min), 200 ml of the reaction mixture was removed and added to a separate sampling tube containing 70 ml 5ϫ Laemmli buffer (40% glycerol, 5% 2
European annals of allergy and clinical immunology, 2018
The aim of this work was to study the effect of industrial processing on the allergenicity of three commonly consumed Moroccan fish species in Fez region (sardine, common pandora, and shrimp). Methods. This work was conducted by a sera-bank obtained from 1248 patients recruited from Fez Hospitals. Their sera were analyzed for specific IgE binding to raw fish extracts. Among them, 60 patients with higher specific IgE levels were selected, and used to estimate the binding variation of IgE to these products under several processing (frying, cooking, canning, marinade, and fermentation) using ELISA analysis. Results. ELISA results demonstrated that all the studied processing cause a reduction in the immunoreactivity of human IgE to fish products, with a high action with marinade and fermentation compared to other processing. This alteration was also observed with rabbit IgG in all processed products, showing that the maximum reduction was marked in fermented sardine with 64.5%, in cooked common pandora with 58%, and in fermented shrimp with 69.2%. Conclusion. In conclusion, our study has shown that the allergenicity of the three studied fish could be reduced by different industrial processes with different degrees.
Clinical and Translational Allergy, 2014
Background: Fish is an important cause of food allergy. Studies on fish allergy are scarce and in most cases limited to serological evaluation. Our objective was to study patterns of self-reported allergy and tolerance to different commonly consumed fish species and its correlation to IgE sensitization to the same species. Methods: Thirty-eight adult fish allergic patients completed a questionnaire regarding atopy, age of onset and symptoms to 13 commonly consumed fish species in the Netherlands (pangasius, cod, herring, eel, hake, pollock, mackerel, tilapia, salmon, sardine, tuna, plaice and swordfish). Specific IgE to these fish extracts were analyzed by ImmunoCAP.
Seafood allergy: A comprehensive review of fish and shellfish allergens
Molecular immunology, 2018
Seafood refers to several distinct groups of edible aquatic animals including fish, crustacean, and mollusc. The two invertebrate groups of crustacean and mollusc are, for culinary reasons, often combined as shellfish but belong to two very different phyla. The evolutionary and taxonomic diversity of the various consumed seafood species poses a challenge in the identification and characterisation of the major and minor allergens critical for reliable diagnostics and therapeutic treatments. Many allergenic proteins are very different between these groups; however, some pan-allergens, including parvalbumin, tropomyosin and arginine kinase, seem to induce immunological and clinical cross-reactivity. This extensive review details the advances in the bio-molecular characterisation of 20 allergenic proteins within the three distinct seafood groups; fish, crustacean and molluscs. Furthermore, the structural and biochemical properties of the major allergens are described to highlight the im...
Food Research International, 2019
Prawn allergy is one of the most common food-borne allergies and current prevention is by avoidance. This review paper summarised different methodologies for the extraction, identification and quantification of prawn protein allergens, reported in various research studies. Following extraction, allergenic components have been analysed using well-established methodologies, such as SDS-PAGE, Immunoblotting, ELISA, CD Spectroscopy, HPLC, DBPCFC, SPT etc. Moreover, the preference towards Aptamer-based technique for allergenicity analysis has also been highlighted in this review paper. The summary of these methodologies will provide a reference platform for present and future research directions.
Indian journal of experimental biology, 2005
Enzymed-linked immunosorbent assay of hilsa and pomfret muscle extracts showed specific IgE binding to ten allergic patients' sera, the results corroborated to that of skin prick test. Comparison of allergen profiles of the two fish extracts by immunoblotting revealed a common antigenic protein of 50 kDa and some high molecular weight fish allergens instead of low molecular weight parvalbumin found in several fishes. Purified and well characterized fish allergens are always considered better than crude fish extracts for diagnostic use.