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BAL ARISI ZEHRİNİN KARAKTERİZASYONUNDA SDS-PAGE ELEKTROFOREZ KULLANILABİLİRLİĞİNİN ARAŞTIRILMASI

Year 2016, Volume: 16 Issue: 2, 49 - 56, 28.04.2017
https://doi.org/10.31467/uluaricilik.379483

Abstract

Arı zehri (bee venom, apizehir) açık renkte, kokusuz, sıvı bir madde olup, keskin, acı bir tada sahip bir
peptit ve protein karışımıdır. Biyolojik aktif değeri yüksek çok sayıda peptit ve proteinden oluşan arı
zehri apiterapi uygulamaları için çok değerlidir. Kozmetikten apiterapi uygulamalarına kadar çok
sayıda kullanım alanı bulan arı zehrinin kullanılmadan önce kalitesinin test edilmesi gerekir. Arı zehri
analizi HPLC, LC-MS/MS, MALDİ-TOF gibi ileri analiz yöntemleri ve laboratuvar donanımı
gerektirmektedir. Yapılan bu çalışmada, arı zehrinin pratik ve ucuz yolla teşhis edilmesine imkân
sağlayan metot geliştirildi. Taze toplanan arı zehrinin ham protein miktarı ile sodyum dodesil sülfatpoliakrilamid
jel (SDS-PAGE) elektroforez kromatografisi yapıldı. Toplam protein miktarı %66 olarak
bulunan arı zehrinin SDS-PAGE ile üç önemli protein bandı elde edildi. Molekül ağırlıkları yaklaşık 30
ve 40 kDa iki küçük ve molekül ağırlığı yaklaşık 4 kDa olan büyük bir protein bandı elde edildi. Arı
zehrinin yaklaşık %60’ını oluşturan melittin küçük molekül kütlesinden dolayı en uzağa yürüyen bant
olup (4 kDa.dan küçük) molekül ağırlığı 30-40 kDa arasında olan iki küçük bantın hyaluronidaz,
fosfolipaz A2 enzimlerine ait oluğu tespit edildi. Sonuç olarak, SDS-PAGE elektroforez tekniği arı
zehrinin pratik olarak teşhis edilmesine uygun bir yöntem olduğu görülmektedir.

References

  • Anonim, 1989. Ari Zehiri Tasarisi. Türk Standartları Enstitüsü. Ankara.
  • Atayoğlu, T. 2012. Apiterapi Açısından Arı Ürünlerinin Kalite Kriterleri Ve Standardizasyonu, Amerikan Hastanesi, Türk Standartları Enstitüsü.
  • Billingham, M. E. J., Morley, J., Hanson, J. M., Shipolini, R. A., Vernon, C. A. 1973. An anti-inflammatory peptide from bee venom. Nature Publishing Group.
  • Bogdanov, S., 2016. Bee Venom: production, composition and quality. In: The bee venom Book, Chapter 1, Muehlethurnen, Switzerland. www.bee-hexagon.net.
  • Bradford, M. M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical biochemistry, 72,1-2, 248-254.
  • Chen J. ve Lariviere W.R. 2010. The nociceptive and anti-nociceptive effects of bee venom injection and therapy: A double-edged sword. Progress in Neurobiology 92,151–183.
  • Danneels, E. L., Van Vaerenbergh, M., Debyser, G., Devreese, B., de Graaf, D. C. (2015). Honeybee venom proteome profile of queens and winter bees as determined by a mass spectrometric approach. Toxins, 7(11), 4468-4483.
  • Gajski, G., Čimbora‐Zovko, T., Rak, S., Rožman, M., Osmak, M., Garaj‐Vrhovac, V. 2014. Combined antitumor effects of bee venom and cisplatin on human cervical and laryngeal carcinoma cells and their drug resistant sublines. Journal of Applied Toxicology, 34,12: 1332-1341.
  • Golden, D.B., Marsh, D.G., Freidhoff, L.R. 1997. Natural history of Hymenoptera Venom Sensitivity in Adults. Journal Allergy Clin Immunology, 100:760-6.
  • Gökmen, E.N. 2008. Venom Biyokimyası, Türkiye Klinikleri J Allergy-Special Topics, 1(1) :22-25. Kelle, İ. 2007. Apiterapi. Dicle Tıp Dergisi, 34: 311- 315.
  • Kim, C. S., Yates, D. M., & Heaney, P. J. 1997. The layered sodium silicate magadiite: an analog to smectite for benzene sorption from water. Clays and clay minerals, 45(6), 881-885.
  • Kim, J. Y., Lee, W. R., Kim, K. H., An, H. J., Chang, Y. C., Han, S. M., & Park, K. K. 2015. Effects of bee venom against Propionibacterium acnes induced inflammation in human keratinocytes and monocytes. International Journal of Molecular Medicine, 35(6), 1651-1656.
  • King, T.P., Guralnick, M. 2004. Hymenoptera allergens. Clin Allergy Immunology, 18: 339-53.
  • Kokot, Z. J., Matysiak, J. (2009). Simultaneous determination of major constituents of honeybee venom by LC-DAD. Chromatographia, 69 (11-12), 1401-1405.
  • Lee JD, Park HJ, Chae Y, Lim S. 2005. An Overview of Bee Venom Acupuncture in the Treatment of Arthritis. Evid Based Complement Journal of Alternative and Complementary Medicine 2: 79–84.
  • Longsworth, L. G., MacInnes, D. A. (1940). An electrophoretic study of nephrotic sera and urine. The Journal of experimental medicine, 71(1): 77-82.
  • Michener, C.D. 2007. The Bees of the World, 2nd ed. Johns Hopkins University Press, Baltimore, MD. Molan, P. C. 2006. The evidence supporting the use of honey as a wound dressing. Int. J. Low Ekstrem Wounds, 5 40–54 10.1177/1534734605286014.
  • Moolenaar, M, Poorter, RL, van der Toorn, PP, Lenderink, AW, Poortmans, P, Gerardus Egberts, AC. 2006. The effect of honey compared to conventional treatment on healing of radiotherapyinduced skin toxicity in breast cancer patients. Acta Oncol, 45: 623–4.
  • Moreno, M., Giralt, E., 2015. Three valuable peptides from bee and wasp venoms for therapeutic and biotechnological use: melittin, apamin and mastoparan. Toxins, 7: 1126e1150.
  • Özbek, H. 1990. Bal Arısı (Apis Mellifera L.) Zehri, Yüksek lisans Tezi, Atatürk Üniversitesi Ziraat Fakültesi Bitki Koruma Bölümü, Erzurum 18s.
  • Pacakova, V., Štulík, K., Hau, P. T., Jelinek, I., Vinš, I., Sýkora, D. (1995). Comparison of high-performance liquid chromatography and capillary electrophoresis for the determination of some bee venom components. Journal of Chromatography A, 700(1), 187-193.
  • Park, H.J. Lee, S.H., Son, D.J., Oh, K.W., Kim, K.H., Song, H.S., Kim, G.J., G.T. Oh, Yoon, D.Y., Hong, J.T. 2004. Antiarthritic effect of bee venom: inhibition of inflammation mediator generation by suppression of NF-kappa B through interaction with the p50 subunit. Arthritis & Rheumatology 50: 3504-3515. Putz, T., Ramoner, R., Gander, H., Rahm, A., Bartsch, G., Thurnher, M. 2006. Antitumor action and immune activation through cooperation of bee venom secretory phospholipase A2 and phosphatidylinositol-(3,4)-bisphosphate. Cancer Immunology, Immunotherapy. 55: 1374-1383.
  • Rybak-Chmielewska, H., Szczêsna, T. 2004. HPLC Study of Chemical Composition of Honeybee (Apis Mellifera L.) Venom. Journal of Apicultural Science. 48 (2): 187-193.
  • Son, D.J., Lee, J.W., Lee, Y.H., Song, H.S., Lee, C.K., Hong, J.T. 2007. Therapeutic application of anti-arthritis, pain-releasing, and anti-cancer effects of bee venom and its constituent compounds. Pharmacology Therapeutics Journal. 115: 246-270.
  • Stangaciu, S. 2006. What is apitherapy? www.apitherapy.com (Accessed 27.10.06).
  • Szokan G., Horvath J., Almas M., Saftics G., Palocz A. (1994). Liquid Chromatographic Analysis and Separation of Polipeptide Components from Honey Bee Venoms. J. Liquid. Chrom., 17 (16):3333-3349.
  • White, J. 2000. Bites and Stings From Venomous Animals a Global Overview. Ther Drug Monitoring. 22: 65-68.
  • Zenon J. K., Matysiak, J., Urbaniak,B., Dereziński, P. 2011. New CZE-DAD method for honeybee venom analysis and standardization of the product. Analytical and Bioanalytical Chemistry. 399:7, 2487–2494.
  • Zumla ve Lulat, 1989. Honey-a remediy rediscovered. Journal of the royal society of medicine. 82 (7): 384-385
Year 2016, Volume: 16 Issue: 2, 49 - 56, 28.04.2017
https://doi.org/10.31467/uluaricilik.379483

Abstract

References

  • Anonim, 1989. Ari Zehiri Tasarisi. Türk Standartları Enstitüsü. Ankara.
  • Atayoğlu, T. 2012. Apiterapi Açısından Arı Ürünlerinin Kalite Kriterleri Ve Standardizasyonu, Amerikan Hastanesi, Türk Standartları Enstitüsü.
  • Billingham, M. E. J., Morley, J., Hanson, J. M., Shipolini, R. A., Vernon, C. A. 1973. An anti-inflammatory peptide from bee venom. Nature Publishing Group.
  • Bogdanov, S., 2016. Bee Venom: production, composition and quality. In: The bee venom Book, Chapter 1, Muehlethurnen, Switzerland. www.bee-hexagon.net.
  • Bradford, M. M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical biochemistry, 72,1-2, 248-254.
  • Chen J. ve Lariviere W.R. 2010. The nociceptive and anti-nociceptive effects of bee venom injection and therapy: A double-edged sword. Progress in Neurobiology 92,151–183.
  • Danneels, E. L., Van Vaerenbergh, M., Debyser, G., Devreese, B., de Graaf, D. C. (2015). Honeybee venom proteome profile of queens and winter bees as determined by a mass spectrometric approach. Toxins, 7(11), 4468-4483.
  • Gajski, G., Čimbora‐Zovko, T., Rak, S., Rožman, M., Osmak, M., Garaj‐Vrhovac, V. 2014. Combined antitumor effects of bee venom and cisplatin on human cervical and laryngeal carcinoma cells and their drug resistant sublines. Journal of Applied Toxicology, 34,12: 1332-1341.
  • Golden, D.B., Marsh, D.G., Freidhoff, L.R. 1997. Natural history of Hymenoptera Venom Sensitivity in Adults. Journal Allergy Clin Immunology, 100:760-6.
  • Gökmen, E.N. 2008. Venom Biyokimyası, Türkiye Klinikleri J Allergy-Special Topics, 1(1) :22-25. Kelle, İ. 2007. Apiterapi. Dicle Tıp Dergisi, 34: 311- 315.
  • Kim, C. S., Yates, D. M., & Heaney, P. J. 1997. The layered sodium silicate magadiite: an analog to smectite for benzene sorption from water. Clays and clay minerals, 45(6), 881-885.
  • Kim, J. Y., Lee, W. R., Kim, K. H., An, H. J., Chang, Y. C., Han, S. M., & Park, K. K. 2015. Effects of bee venom against Propionibacterium acnes induced inflammation in human keratinocytes and monocytes. International Journal of Molecular Medicine, 35(6), 1651-1656.
  • King, T.P., Guralnick, M. 2004. Hymenoptera allergens. Clin Allergy Immunology, 18: 339-53.
  • Kokot, Z. J., Matysiak, J. (2009). Simultaneous determination of major constituents of honeybee venom by LC-DAD. Chromatographia, 69 (11-12), 1401-1405.
  • Lee JD, Park HJ, Chae Y, Lim S. 2005. An Overview of Bee Venom Acupuncture in the Treatment of Arthritis. Evid Based Complement Journal of Alternative and Complementary Medicine 2: 79–84.
  • Longsworth, L. G., MacInnes, D. A. (1940). An electrophoretic study of nephrotic sera and urine. The Journal of experimental medicine, 71(1): 77-82.
  • Michener, C.D. 2007. The Bees of the World, 2nd ed. Johns Hopkins University Press, Baltimore, MD. Molan, P. C. 2006. The evidence supporting the use of honey as a wound dressing. Int. J. Low Ekstrem Wounds, 5 40–54 10.1177/1534734605286014.
  • Moolenaar, M, Poorter, RL, van der Toorn, PP, Lenderink, AW, Poortmans, P, Gerardus Egberts, AC. 2006. The effect of honey compared to conventional treatment on healing of radiotherapyinduced skin toxicity in breast cancer patients. Acta Oncol, 45: 623–4.
  • Moreno, M., Giralt, E., 2015. Three valuable peptides from bee and wasp venoms for therapeutic and biotechnological use: melittin, apamin and mastoparan. Toxins, 7: 1126e1150.
  • Özbek, H. 1990. Bal Arısı (Apis Mellifera L.) Zehri, Yüksek lisans Tezi, Atatürk Üniversitesi Ziraat Fakültesi Bitki Koruma Bölümü, Erzurum 18s.
  • Pacakova, V., Štulík, K., Hau, P. T., Jelinek, I., Vinš, I., Sýkora, D. (1995). Comparison of high-performance liquid chromatography and capillary electrophoresis for the determination of some bee venom components. Journal of Chromatography A, 700(1), 187-193.
  • Park, H.J. Lee, S.H., Son, D.J., Oh, K.W., Kim, K.H., Song, H.S., Kim, G.J., G.T. Oh, Yoon, D.Y., Hong, J.T. 2004. Antiarthritic effect of bee venom: inhibition of inflammation mediator generation by suppression of NF-kappa B through interaction with the p50 subunit. Arthritis & Rheumatology 50: 3504-3515. Putz, T., Ramoner, R., Gander, H., Rahm, A., Bartsch, G., Thurnher, M. 2006. Antitumor action and immune activation through cooperation of bee venom secretory phospholipase A2 and phosphatidylinositol-(3,4)-bisphosphate. Cancer Immunology, Immunotherapy. 55: 1374-1383.
  • Rybak-Chmielewska, H., Szczêsna, T. 2004. HPLC Study of Chemical Composition of Honeybee (Apis Mellifera L.) Venom. Journal of Apicultural Science. 48 (2): 187-193.
  • Son, D.J., Lee, J.W., Lee, Y.H., Song, H.S., Lee, C.K., Hong, J.T. 2007. Therapeutic application of anti-arthritis, pain-releasing, and anti-cancer effects of bee venom and its constituent compounds. Pharmacology Therapeutics Journal. 115: 246-270.
  • Stangaciu, S. 2006. What is apitherapy? www.apitherapy.com (Accessed 27.10.06).
  • Szokan G., Horvath J., Almas M., Saftics G., Palocz A. (1994). Liquid Chromatographic Analysis and Separation of Polipeptide Components from Honey Bee Venoms. J. Liquid. Chrom., 17 (16):3333-3349.
  • White, J. 2000. Bites and Stings From Venomous Animals a Global Overview. Ther Drug Monitoring. 22: 65-68.
  • Zenon J. K., Matysiak, J., Urbaniak,B., Dereziński, P. 2011. New CZE-DAD method for honeybee venom analysis and standardization of the product. Analytical and Bioanalytical Chemistry. 399:7, 2487–2494.
  • Zumla ve Lulat, 1989. Honey-a remediy rediscovered. Journal of the royal society of medicine. 82 (7): 384-385
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Publication Date April 28, 2017
Acceptance Date January 9, 2017
Published in Issue Year 2016 Volume: 16 Issue: 2

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Vancouver Kolaylı S. BAL ARISI ZEHRİNİN KARAKTERİZASYONUNDA SDS-PAGE ELEKTROFOREZ KULLANILABİLİRLİĞİNİN ARAŞTIRILMASI. U. Arı. D.-U. Bee J. 2017;16(2):49-56.

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