Despite being known as resistant proteins, peanut allergens (Ara h 1 and Ara h 2)
can be digested and cause allergic reactions. Making the allergens more resistant to
digestion may aid in non-absorption and excretion of the allergens. Our objectives
were to make Ara h 1 and Ara h 2 more resistant to digestion and test them in a
model system using trypsin as the digestive enzyme. The resistant allergens were
prepared by covalently attaching p-aminobenzamidine (pABA), a protease inhibitor,
to peanut allergens in an extract or on a PVDF membrane using glutaraldehyde, and
were then tested for resistance to trypsin digestion. SDS-PAGE and Western blot
were performed to determine the allergenic capacity of the modified allergens. A
control was prepared using glycine instead. Results showed that Ara h 2, when covalently
attached with pABA, was more resistant to trypin digestion than the native allergen.
Similarly, Ara h 1, prepared on a PVDF membrane and treated with pABA,
displayed a resistance to trypsin digestion. Treatment of the allergens with glycine (a
control) instead of pABA showed that the modified allergens were as digestible as
native allergens. Blot assays showed that the pABA-treated allergens exhibited a lower
allergenic capacity than native allergens. It was concluded that pABA, when attached
to peanut allergen Ara h 1 or Ara h 2, inhibited digestion of the allergen by
trypsin and reduced their allergenic capacity as well.
References
[1]
Toomer, O.T., Do, A., Pereira, M. and Williams, K. (2013) Effect of Simulated Gastric and Intestinal Digestion on Temporal Stability and Immunoreactivity of Peanut, Almond, and Pine Nut Protein Allergens. Journal of Agricultural and Food Chemistry, 61, 5903-5913. https:/doi.org/10.1021/jf400953q
[2]
Calloway, D.H. and Kretsch, M.J. (1978) Protein and Energy Utilization in Men Given a Rural Quatemalan Diet and Egg Formulas with and without Added Oat Bran. The American Journal of Clinical Nutrition, 31, 1118-1126.
[3]
Southgate, D.A.T. and Durnin, J.V.G.A. (1970) Calorie Conversion Factors. An Experimental Reassessment of the Factors Used in the Calculation of the Energy Value of Human Diets. British Journal of Nutrition, 24, 517-535. https:/doi.org/10.1079/BJN19700050
[4]
Gurr, M.I. (2012) Dietary Fats in Relation to Weight Control. Lipids in Nutrition and Health, 5, 161-179. https:/doi.org/10.1533/9780857097989.161
Burks, A.W., Christie, L., Althage, K.A., Kesler, J.M. and Allgoody, G.S. (2001) Randomized, Double-Blind, Placebo-Controlled, Food Allergy Challenge to Olestra Snacks. Regulatory Toxicology and Pharmacology, 3, 4178-4181.
[7]
Vinson, J.A., Kharrat, H.A.I. and Shuta, D. (2009) Investigation of an Amylase Inhibitor on Human Glucose Absorption after Starch Consumption. Open Nutraceuticals Journal, 2, 88-91. https:/doi.org/10.2174/1876396000902010088
[8]
Udani, J., Hardy, M. and Madsen, D.C. (2004) Blocking Carbohydrate Absorption and Weight Loss: A Clinical Trial Using Phase 2 Brand Proprietary Fractional White Bean Extract. Alternative Medicine Review, 9, 63-69.
[9]
Celleno, L., Tolaini M.V., D’Amore, A., Perricone, N.V. and Preuss, H.G. (2007) A Dietary Supplement Containing Standardized Phaseolus Vulgaris Extract Influences Body Composition of Overweight Men and Women. International Journal of Medical Sciences, 4, 45-52. https:/doi.org/10.7150/ijms.4.45
[10]
Zhao, W., Iyer, V., Flores, F.P., Donhowe, E. and Kong, F. (2013) Microencapsulation of Tannic Acid for Oral Administration to Inhibit Carbohydrate Digestion in the Gastrointestinal Tract. Food Function, 4, 899-900. https:/doi.org/10.1039/c3fo30374h
[11]
Stojadinovic, M., Radosavljevic, J., Ognjenovic, J., Vesic, J., Prodic, I., Stanic-Vucinic, D. and Cirkovic Velickovic, T. (2013) Binding Affinity between Dietary Polyphenols and β-Lactoglobulin Negatively Correlates with the Protein Susceptibility to Digestion and Total Antioxidant Activity of Complexes Formed. Food Chemistry, 136, 1263-1271. https:/doi.org/10.1016/j.foodchem.2012.09.040
[12]
Sudhair, J., Pushparaj, T., Thavarajah, D., Premakumara, S., Abeysekara, K. and Sotheeswaran, S. (2015) Rice (Oryza sativa L.) Resistant Starch and Novel Processing Methods to Increase Resistant Starch Concentration. American Chemical Society National Meeting, Denver, AGFD, 33.
[13]
White, B.L., Shi, X., Burk, C.M., Kulis, M., Burks, A.W., Sanders, T.H. and Davis, J.P. (2014) Strategies to Mitigate Peanut Allergy: Production, Processing, Utilization, and Immunotherapy Considerations. Annual Review of Food Science and Technology, 5, 155-176. https:/doi.org/10.1146/annurev-food-030713-092443
[14]
Chung, S.Y. and Reed, S. (2014) Reducing Food Allergy: Is There Promise for Food Applications? Current Pharmaceutical Design, 20, 924-930. https:/doi.org/10.2174/13816128113199990043
[15]
Zhao, X., Yang, W., Chung, S.Y., Sims, C.A., Otwell, S.W. and Rababah, T.M. (2014) Reduction of IgE Immunoreactivity of Whole Peanut (Arachis hypogaea L.) after Pulsed Light Illumination. Food and Bioprocess Technology, 7, 2637-2645. https:/doi.org/10.1007/s11947-014-1260-7
[16]
Chung, S.Y. and Reed, S. (2015) IgE Binding to Peanut Allergens Is Inhibited by Combined D-Aspartic and D-Glutamic Acids. Food Chemistry, 166, 248-253. https:/doi.org/10.1016/j.foodchem.2014.06.004
[17]
Chung, S.Y., Mattison, C.P., Reed, S., Wasserman, R.L. and Desormeaux, W.A. (2015) Treatment with Oleic Acid Reduces IgE Binding to Peanut and Cashew Allergens. Food Chemistry, 180, 295-300. https:/doi.org/10.1016/j.foodchem.2015.02.056
[18]
Fasoli, E., Reyes, Y.R., Guzman, O.M., Rosado, A., Cruz, V.R., Borges, A., Martinez, E. and Bansal, V. (2013) Para-Aminobenzamidine Linked Regenerated Cellulose Membranes for Plasminogen Activator Purification: Effect of Spacer Arm Length and Ligand Density. Journal of Chromatography B, 930, 13-21. https:/doi.org/10.1016/j.jchromb.2013.04.025
[19]
Evans, S.A., Olson, S.T., and Shore, J.D. (1982) p-Aminobenzamidine as a Fluorescent Probe for the Active Site of Serine Proteases. Journal of Biological Chemistry, 257, 3014-3017.
[20]
Fernandez, J., Andrews, L. and Mische, S.M. (1994) A One-Step Enzymatic Digestion Procedure for PVDF-Bound Proteins That Does Not Require PVP-40. Techniques in Protein Chemistry, 5, 215-222. https:/doi.org/10.1016/B978-0-12-194710-1.50029-1
[21]
Vestling, M.M. and Fenselau, C.C. (1994) Protease Digestion on PVDF Membrane for Matrix-Assisted Laser Desorption Mass Spectrometry. Techniques in Protein Chemistry, 5, 59- 67. https:/doi.org/10.1016/B978-0-12-194710-1.50012-6
[22]
Lehmann, K., Schweimer, K., Reese, G., Randow, S., Suhr, M. and Becker, W.M. (2006) Structure and Stability of 2S Albumin-Type Peanut Allergens: Implications for the Severity of Peanut Allergic Reactions. Biochemical Journal, 395, 463-472. https:/doi.org/10.1042/BJ20051728
[23]
Shi, X., Guo, R., White, B.L., Yancey, A., Sanders, T.H. and Davis, J.P. (2013) Allergenic Properties of Enzymatically Hydrolyzed Peanut Flour Extracts. International Archives of Allergy and Immunology, 162, 123-130. https:/doi.org/10.1159/000351920
[24]
Chung, S.Y. and Reed S. (2012) Removing Peanut Allergens by Tannic Acid. Food Chemistry, 134, 1468-1473. https:/doi.org/10.1016/j.foodchem.2012.03.057
[25]
Hazebrouck, S., Guillon, B., Drumare, M.F., Paty, E., Jean-MichelWal, J.M. and Bernard, H. (2012) Trypsin Resistance of the Major Peanut Allergen Ara h 6 and Allergenicity of the Digestion Products Are Abolished after Selective Disruption of Disulfide Bonds. Molecular Nutrition and Food Research, 56, 548-557. https:/doi.org/10.1002/mnfr.201100614