First purification of glutathione S-transferase enzyme from honey bees (Apis mellifera L.) and effects of heavy metal ions on bioactivity

Öz

Honey bees (Apis mellifera L.) in the Bingöl province of Türkiye play a crucial role in both pollination and contributing to biological processes through their bioactive compounds. Owing to these functions, they possess anticancer, anti-inflammatory, ecological, and economic significance. Glutathione S-transferases (GSTs; EC 2.5.1.18) are detoxification enzymes found in all cellular organisms, including bacteria, plants, insects, and humans. Purifying the glutathione S-transferase enzyme (GST; EC 2.5.1.18) from Apis mellifera L. and examining how different metal ions affect the activity of the purified enzyme were the objectives of this study. The purification process was performed in two steps: homogenate preparation and glutathione-agarose affinity chromatography. The GST was purified with a specific activity of 0.379 EUmg-1 protein, yielding a 7.7% yield and a 77.2-fold purification. SDS-PAGE analysis was used to verify the purity of the enzyme, and only a single protein band was detected. Metal ion effects were evaluated through activity percentage plots, inhibition constants (Ki), and half-maximal inhibitory concentration (IC₅₀) values. Inhibition types were determined using Lineweaver–Burk plots for the metal ions exhibiting inhibitory effects. Pb²⁺ had no significant effect on enzyme activity. In contrast, Fe²⁺ and Co²⁺ ions activated the enzyme, while Hg²⁺, Ag⁺, and Cu²⁺ showed non-competitive inhibition. To the best of our knowledge, this is the first report on the purification of GST from honey bees.

Anahtar Kelimeler

• Bee
• Glutathione S-transferase
• Purification
• Metal ions

Destekleyen Kurum

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Proje Numarası

Bu araştırma kamu, ticari veya kâr amacı gütmeyen sektörlerdeki fon sağlayıcı kuruluşlardan herhangi bir özel hibe almamıştır.

Etik Beyan

This study does not contain any experiments with human participants or animals

Bal arılarından (Apis mellifera L.) glutatyon S-transferaz enziminin ilk saflaştırılması ve ağır metal iyonlarının biyoaktivite üzerine etkileri

Öz

Bingöl ilindeki bal arıları (Apis mellifera L.) hem çiçeklerin tozlaşmasını sağlar hem de taşıdıkları önemli biyoaktif bileşenlerle biyolojik fonksiyonlara katkıda bulunurlar. Bu önemli rollerinden dolayı antikanser, antiinflamatuar, ekolojik ve ekonomik öneme sahiptirler. Glutatyon transferazlar (EC 2.5.1.18), tüm hücresel organizmalarda (bakteriler, bitkiler, böcekler ve insanlar) bulunan detoksifikasyon enzimleridir. Bu çalışmada ilk kez arı dokularından (Apis mellifera L.) glutatyon S-transferaz enziminin (GST; EC 2.5.1.18) saflaştırılması ve bazı metal iyonlarının enzim aktivitesi üzerine etkilerinin araştırılması hedeflenmiştir. Saflaştırma işlemi, homojenat hazırlama ve glutatyon-agaroz afinite kromatografisi olmak üzere iki adımda gerçekleştirildi. Çalışmada, arı dokularından 0,379 EUmg-1 protein spesifik aktivitesine sahip glutatyon S-transferaz enzimi; % 7,7 verimle 77,2 kat saflaştırıldı. Enzimin saflığı SDS-PAGE ile kontrol edildi ve tek bant elde edildi. Ayrıca bazı metal iyonlarının enzim aktivitesi üzerine etkileri incelendi. Metal iyonları için % activite - [metal iyonu] ve Lineweaver-Burk grafikleri yardımıyla sırası ile IC50 değerleri ve Ki sabitleri hesaplandı ve inhibitör etki gösteren metal iyonları için inhibisyon tipleri belirlendi. Metal iyonlarından Pb+2 iyonu enzim aktivitesi üzerinde herhangi bir etki oluşturmazken, Fe+2, Co+2 iyonları enzimi aktive, Hg+2, Ag+, Cu+2 iyonlarının enzimi yarışmasız olarak inhibe ettikleri tespit edildi.

Anahtar Kelimeler

• Arı
• Glutatyon S-transferaz
• Saflaştırma
• Metal iyonlar

Destekleyen Kurum

Bu araştırma kamu, ticari veya kâr amacı gütmeyen sektörlerdeki fon sağlayıcı kuruluşlardan herhangi bir özel hibe almamıştır.

Proje Numarası

Bu araştırma kamu, ticari veya kâr amacı gütmeyen sektörlerdeki fon sağlayıcı kuruluşlardan herhangi bir özel hibe almamıştır.

Etik Beyan

Bu çalışma insan katılımcıları veya hayvanlarla herhangi bir deney içermemektedir

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