Extraction and Purification of the Potential Allergen Proteins from Aspergillus Fumigatus

Abstract

Allergic diseases are an important public health problem today and are increasing rapidly around the world. According to the research conducted by the World Allergy Organization (WAO), %22 of the world population is struggling with allergic diseases. In our country, one out of every five citizens complains of at least one form of allergy, and it is the third most common chronic disease in children. Some of the allergic diseases develop suddenly and are life threatening. The diagnosis and treatment of allergic diseases is one of the most expensive diseases in our country and in the world. It is triggered by the production of IgE(Immunoglobulin E) in the body against antigens in individuals sensitive to allergen substances. Allergens that play a role in the development of allergic diseases are protein molecules that stimulate IgE synthesis. Pollen, fungus and house dust mites are the most common allergens due to their widespread presence in nature. The presence of fungal spores in the atmosphere for a long time with the effect of air flow increases the possibility of people coming into contact with these allergens. It is very important for health to determine which factors individuals have allergies to. Among the current diagnostic methods; respiratory function tests, skin tests with allergens (prick tests, interdermal tests, patch tests), serum total IgE determination and procovation tests. All of the existing tests used in the identification of allergic diseases in our country are of foreign origin and are imported. One of the fungi that cause allergies is Aspergillus fumigatus from the genus Aspergillus. In our study, we aimed to purify Aspergillus fumigatus allergen proteins and produce allergens that can be used in allergy diagnosis methods. For this purpose, protein isolation and purification of Aspergillus fumigatus, which was cultured and multiplied in SDA (Sabouraud Dextrose Agar) medium in the laboratory, was performed with 2 different methods. The protein amount of the product obtained was determined by the BCA(Bicinchoninic Acid) method. In our studies, it was determined that the protein content of the extract prepared with chloroform/methanol solution was higher than the protein amount of the extract prepared with ethanol solution.

Keywords

• Allergy
• Aspergillus fumigatus
• Allergen protein
• Fungal allergy
• BCA (Bicinchoninic Acid)

References

1. Asan, A., Sen, B., & Sarica, S. (2002). Airborne fungi in urban air of Edirne city (Turkey). Section Botany, 57(1), 59-68.
2. Klironomos, J. N., Rillig, M. C., Allen, M. F., Zak, D. R., Pregitzer, K. S., & Kubiske, M. E. (1997). Increased levels of airborne fungal spores in response to Populus tremuloides grown under elevated atmospheric CO2. Canadian Journal of Botany, 75(10), 1670-1673.
3. Corsico, R., Cinti, B., Feliziani, V., Gallesio, M. T., Liccardi, G., Loreti, A., ... & Zerboni, R. (1998). Prevalence of sensitization to Alternaria in allergic patients in Italy. Annals of Allergy, Asthma & Immunology, 80(1), 71-76.
4. Guneser S, Atici A, Koksal F, & Yaman A. (1994). Mold allergy in Adana, Turkey. Allergol Immunopathol 22(2), 52-4.
5. Yurdakulol, E., & Cansaran, D. (2004). Tohumsuz bitkiler I laboratuvar kılavuzu. 74, Basılmamış.
6. Durugbo, E. U., Kajero, A. O., Omoregie, E. I., & Oyejide, N. E. (2013). A survey of outdoor and indoor airborne fungal spora in the Redemption City, Ogun State, south-western Nigeria. Aerobiologia, 29(2), 201-216. Kurup, V. P., Banerjee, B., Hemmann, S., Greenberger, P. A., Blaser, K., & Crameri, R. (2000). Selected recombinant Aspergillus fumigatus allergens bind specifically to IgE in ABPA. Clinical and experimental allergy: journal of the British Society for Allergy and Clinical Immunology, 30(7), 988-993.
7. Latgé, J. P. (2001). The pathobiology of Aspergillus fumigatus. Trends in microbiology, 9(8), 382-389.
8. Walker, J. M. (2002). SDS polyacrylamide gel electrophoresis of proteins. In The protein protocols handbook (pp. 61-67). Humana Press.

9. Smith, P. E., Krohn, R. I., Hermanson, G. T., Mallia, A. K., Gartner, F. H., Provenzano, M., ... & Klenk, D. C. (1985). Measurement of protein using bicinchoninic acid. Analytical biochemistry, 150(1), 76-85. Kahlert, S. (1996). Identification and characterisation of a specific extracellular component of Aspergillus fumigatus. [Doctoral dissertation], University of Surrey.