Bal arısı zehrinin karakterizasyonunda sds-page elektroforez kullanılabilirliğinin araştırılması
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Date
2017-01-09
Authors
Şirin, Yakup
Çakır, Hilal Ebru
Can, Zehra
Yıldız, Oktay
Kolaylı, Sevgi
Journal Title
Journal ISSN
Volume Title
Publisher
Uludağ Üniversitesi
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.
Apitoxin called or bee venom is light color, odor-free, liquid, its taste is bitter and apitoxin is a protein mixture. Peptites and protein of the bee venom have many biological activities and its very important for apitherapic applications. Bee venom is used in many areas like apitherapeutic applications and cosmetics, and should be analyzed before uses. Bee venom analyzed need sophisticated analyzed techniques, such as HPLC, LC-MS/MS, MALDI-TOF and developed equipment. In this study, an identification method of bee venoms was developed, and this method provides us to identify bee venoms in a cheap and simple way. Crude protein amont and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the fresh collected bee venom was analysed. Total protein content of the sample was 66% and three important protein bands was obtained. Two small bands of nearly 30 and 40 kDa and a big band of nearly 4 kDa was obtained of the SDS-PAGE. The smallest band (4 kDa) represents melittin, which forms approximately %60 of dried bee venom, because of the small molecular mass of melittin. Also, the other bands (30 and 40 kDa) represent hyaluronidase and phospholipase A2, which are characteristic proteins of bee venom. In conclusion, SDS-PAGE gel electrophoresis is very suitable for determination identification bee venom that was a practical way.
Apitoxin called or bee venom is light color, odor-free, liquid, its taste is bitter and apitoxin is a protein mixture. Peptites and protein of the bee venom have many biological activities and its very important for apitherapic applications. Bee venom is used in many areas like apitherapeutic applications and cosmetics, and should be analyzed before uses. Bee venom analyzed need sophisticated analyzed techniques, such as HPLC, LC-MS/MS, MALDI-TOF and developed equipment. In this study, an identification method of bee venoms was developed, and this method provides us to identify bee venoms in a cheap and simple way. Crude protein amont and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the fresh collected bee venom was analysed. Total protein content of the sample was 66% and three important protein bands was obtained. Two small bands of nearly 30 and 40 kDa and a big band of nearly 4 kDa was obtained of the SDS-PAGE. The smallest band (4 kDa) represents melittin, which forms approximately %60 of dried bee venom, because of the small molecular mass of melittin. Also, the other bands (30 and 40 kDa) represent hyaluronidase and phospholipase A2, which are characteristic proteins of bee venom. In conclusion, SDS-PAGE gel electrophoresis is very suitable for determination identification bee venom that was a practical way.
Description
Keywords
Arı zehri, SDS-PAGE elektroforez, Melittin, Bee venom, SDS-PAGE electrophoresis
Citation
Şirin, Y. vd. (2016). ''Bal arısı zehrinin karakterizasyonunda sds-page elektroforez kullanılabilirliğinin araştırılması''. Uludağ Arıcılık Dergisi, 16(2), 49-56.