Effect of Heat Treatment and Enzymatic Hydrolysis on Reduction in Allergenicity of Milk Proteins

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β -lactoglobulin, allergenicity, Ig E, ELISA, enzyme hydrolysis, heat treatment
Dairy Chemistry


The allergy due to different fractions of milk protein was estimated by the level of milk specific Ig E blood sera of children suspected for milk allergy with a history of symptoms such as wheezing, diarrhoea or dermatitis. It was observed that among the milk proteins β - lactoglobulin of whey protein is the most potent allergen. Effect of heat treatment and enzymatic hydrolysis on reduction in β - lactoglobulin allergenicity was investigated by inhibition ELISA while heat treatment could reduce the allergenicity only to the extent of 12.5%, enzymatic hydrolysis could reduce the allergenicity significantly. The extent of hydrolysis as well as the enzyme used had significant role in reduction of milk protein allergenicity. The degree of hydrolysis corresponded well with the reduction of milk protein allergenicity. Though chymotrypsin was effective in hydrolyzing β - lactoglobulin, neutrase was more effective in reducing the allergenicity by 52% even at lower degree of hydrolysis. The combination of chymotrypsin and neutrase was found to be more useful in hydrolysis as well as reduction in allergenicity of the protein.


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How to Cite

Pushpa, B., Bhat, G., & Jayaprakasha, H. (2018). Effect of Heat Treatment and Enzymatic Hydrolysis on Reduction in Allergenicity of Milk Proteins. The Indian Journal of Nutrition and Dietetics, 55(2), 156–165. https://doi.org/10.21048/ijnd.2018.55.2.19651



Original Articles



Koletzko, S., Niggemann, Arato, A., Dias, J.A. and Heuschkel, R. Diagnosis approach and management of cow's milk protein allergy in infants and children. Espghan GI committee practical Guidelines. J.Pediat. Gastroenterol. Nutr., 2012, 55, 221-229.

Calatayud, M.C., Muriel Garcí­a, A., Martorell Aragonés, A. and De La Hoz Caballer, B.

Safety and efficacy profile and immunological changes associated with oral immunotherapy for IgE-mediated cow's milk allergy in children: systematic review and meta-analysis. J. Invest. Allergol. Clin. Immunol., 2014, 24, 298-307.

Boyle, R.J., Ierodiakonou, D., Khan, T., Chivinge, J., Robinson, Z., Geoghegan, N., Jarrold, K., Afxentiou, T., Reeves, T., Cunha, S., Trivella, M., Garcia-Larsen, V. and Leonardi-Bee, J. Hydrolysed formula and risk of allergic or autoimmune disease: systematic review and meta-analysis. Brit. Med. J., 2016, 352.

Cuomo, B., Indirli, G.C., Bianchi, A., Arasi, S., Caimmi, D., Dondi, A., La Grutta, S., Panetta, V., Verga, M.C and Calvani, M. Specific IgE and skin prick tests to diagnose allergy to fresh and baked cow's milk according to age: a systematic review. Ital. J. Pediat., 2017, 43, 93.

Taylor, S.L. Food allergies. Fd. Technol., 1985, 39, 98-105.

Host, A. Frequency of cow's milk allergy in childhood. Ann. Allergy Asthma Immunol., 2002, 89, 33-37.

Rona. R.J., Keil, T. and Summan, C. The prevalence of food allergy: A meta analysis. J. Allergy Clin. Immunol., 2007, 120, 138-146.

Lifschitz, C and Szajewska, H. Cow's milk allergy: evidence-based diagnosis and management for the practitioner. Eur. J. Pediat., 2015, 174, 141-150.

Bianca, M.E., Gesinne, M.T. and Yuan, V. Preventive possibilities within the context of cow's milk allergy. ACI Intern., 2000, 12, 68-75.

Brill, L.H. Approach to milk protein allergy in infants. Canabian Family Physian. Official Publication of the College of Family Physian of Canada., 2008, 54, 1258-1264.

Brozek, J. and Schunemann, H. World Allergy Organization (WAO) diagnosis and rationale for action against cow's milk allergy guidelines. Pediat. Allergy Immunol., 2010, 21, 121-125.

Martorell-Aragonés, A., Echeverrí­a-Zudaire, L., Alonso-Lebrero, E., Boné-Calvo, J., Martí­nMuñoz, M.F. and Nevot-Falcó, S. Position document: IgE-mediated cow's milk allergy. Allergol. Immunopathol., (Madr) (Practice Guideline. Review), 2015, 43, 507-526.

Adolpson, C.A., Gleich, G.J. and Yunginger, J.W. Standardization of Allergens. In: Manual of Clinical Immunology. 3rd Edn. Washington DC., 1986, 652-659.

Vila, L., Beyer. K., Javviren, K.M., Chatchatte, P., Bardira, L. and Sampson, H.A. Role of conformational and linear epitops in the achievement of tolerance in cow's milk allergy. Clin. Expt. Allergy, 2001, 31, 1599-1606.

Bernard, H., Creminon, C., Yvonm, M. and Wal, J.M. Specificity of the human IgE response to the different purified caseins in allergy to cow's milk proteins. Allergy, 2008, 63, 198-204.

Schulveister, V., Hochwallner, H. and Swoboda. Cloning, expression and mapping of allergenic determinants of αs1-casein, a major cow's milk allergen. J. Immunol., 2009, 182, 7019-7029.

Otani, H., Morita, S. and Tokita, F. Ig M and Ig G antibody responses of rabbits to cow αs1-casein estimated on the basis of peptide fragments derived from αs1-casein. Japanese J. Zootech. Sci., 1985, 341-356.

Hochwallner, H., Schulmeister, U. and Swoboda, I., Spitzauer, S. and Valenta, R. Cow's milk allergy: From allergens to new forms of diagnosis, therapy and prevention. Methods, 2014, 66, 22-33.

Giampietro, P.G., Kjellman, N.I., Oldaeus, G., Wouters-Wesseling, W. and Businco, L.

Hypoallergenicity of an extensively hydrolyzed whey formula. Pediat. Allergy Immunol., 2001, 12, 83-86.

Aas, K. Fish allergy and the codfish allergen model. In : Brostoff, J. and Challacombe, S.J.

Edn. Food allergy and Intolerance. Bailliere Tindall, London, UK, 1987, 356-366.

Thompson, M.P. and Kiddy, C.A. Genetic polymorphism in caseins of cow's milk-III. Isolation and properties of αs1 - caseins A, B and C. J. Dairy Sci., 1964, 47, 633-637.

Hipp, N.J., Groves, M.L., Custa, J.M. and Mameekin, T.L. Separation of αs1, β and γ - casein. J. Dairy Sci., 1952, 35, 272.

Nissen, J.A. Enzymatic hydrolysis of proteins for increased solubility. J. Agric. Fd. Chem., 1986, 24, 16.

Nakamura, T., Sado, H. and Syukunobe, Y. Production of low antigenic whey protein hydrolysates by enzymatic hydrolysis. Milchwissenchaft., 1993, 48, 141-144.

Plebani, A., Ugazio, A.G., Avanzini, A.M., Monafo, V. and Robertto, B.G. An enzyme linked immune sorbent assay for cow's milk protein specific Ig E using biotinylated antigen. J. Immunol. Methods, 1986, 90, 241-246.

Host, A., Halken, S. and Jacobsen, H.P. Clinical course of cow's milk protein allergy / intolerance and atopic diseases in childhood. Pediat. Allergy Immunol., 2002, 13, 23-28.

Schmidt, D.T. and Poll, J.K. Enzymatic hydrolysis of whey protein. Netherlands Milk Dairy J., 1995, 45, 225-240.28. Selo, A., Wauer, W.J. and Bak, H.J. The importance of peptide lengths in hypoallergenic infant formulae. Trends in Fd. Sci.Technol., 1999, 4, 16-21.

Bahna, S.L. and Gandhi, M.D. Milk Hypersensitivity. I. Pathogenisis and symptomatology. Ann. Allergy., 1983, 50, 218-223.

Wal, J.M. Structure and function of milk allergens. Allergy, 2001, 56, 35-38.

Adams, S.L., Barmen, D., Walsh, B.J., Pearct, P.J., Hill, D.J. and Howden, M.E. Human Ig E binding synthetic peptide of bovine β-lactoglobulin and α - lactalbumin. In vitro crossreactivity of the allergens. Immunol. Cell Biol., 1991, 69, 191-197.

Fiocchi, A., Brozek, J., Schünemann, H., Bahna, S.L., von Berg, A., Beyer, K., Bozzola, M. and Bradsher. World Allergy Organization (WAO) Diagnosis and Rationale for Action against Cow's Milk Allergy (DRACMA) Guidelines. J. Pediat. Allergy Immunol., 2010, 21, 121-125.

Dupont, C., Soulaina, P. and Lapillinne, A. Atopy patch test for early diagnosis of cow's milk allergy in preterm infants. J. Pediat. Gastroenterol. Nutr., 2010, 50, 463-464.

Sovage, J. and Christina, B. and Johns, B.A. Food allergy: Epidemiology and natural history. Immunol. Allergy Clin. North Am., 2015, 35, 45 - 59.

Vandenplas, Y. Prevention and management of cow's milk allergy in non-exclusively breastfed infants. Nutr., 2017, 9, 731.