Polinükleer Metal Komplekslerin Plazmid DNA, Alkalen Proteaz Üzerine Etkileri ve Antioksidan Kapasiteleri

Abstract

Bu çalışmada, daha önce sentezlenmiş olan ,N′-Bis[1-(4-fenilfenil)-2-hidroksiimino-2-(pirolidino)-1-etiliden]-1,3-propandiamin ve bunun heterodinükleer Cu(II)-Mn(II) ve Cu(II)-Co(II) kompleksileri, N,N′-Bis[1-(4-fenilfenil)-2-hidroksiimino-2-(4-kloroanilino)-1-etiliden]-1,3-propandiamin ve bunun homodinükleer Cu(II) kompleksi ve homotrinükleer Cu(II) komplekslerinin Bacillus subtilis kaynaklı alkalen proteaz ve plazmid DNA’ya olan etkileri ile antioksidan aktiviteleri araştırılmıştır. Enzim, uygun koşullarda üretildikten sonra amonyum sülfat çöktürme yöntemi ve jel filtrasyon tekniği ile %9,9 verimle ve 69,6 kat saflaştırılmış; SDS-PAGE ile protein molekülünün ağırlığı 30,0 kDa bulunmuştur. Komplekslerin proteaz aktivitesini artırdığı belirlenmiştir. Antioksidan testlerde, homodinükleer Cu(II) içeren B2 kompleksi en yüksek etkiyi göstermiştir. Ayrıca, Cu(II) komplekslerinin DNA’nın doğrusal formunu artırdığı, H₂O₂ varlığında bu etkinlik belirgin şekilde artmıştır. Komplekslerin bazıları tamamen DNA’yı parçalarken bazıları ise hidrojen peroksit içeriğiyle süper sarmal şeklini tümüyle parçalamıştır.

Keywords

• Alkaline proteaz
• Bacillus subtilis
• Schiff bazı
• Antioksidan
• Plazmid DNA.

Supporting Institution

Süleyman Demirel Üniversitesi Bilimsel Araştırma Projeleri Yönetim Birimi

Project Number

2146-D-10

Ethical Statement

Makalede 2020 yılı öncesi araştırma verileri kullanılmıştır ve doktora tezine ait çalışmalardan üretilmiştir. Bu çalışmada, “Yükseköğretim Kurumları Bilimsel Araştırma ve Yayın Etiği Yönergesi” kapsamında uyulması gerekli tüm kurallara uyulduğunu, bahsi geçen yönergenin “Bilimsel Araştırma ve Yayın Etiğine Aykırı Eylemler” başlığı altında belirtilen eylemlerden hiçbirinin gerçekleştirilmediğini taahhüt ederiz.

Thanks

Süleyman Demirel Üniversitesi Bilimsel Araştırma Projeleri Yönetim Birimi’ne ve Süleyman Demirel Üniversitesi Fen-Edebiyat Fakültesi Kimya Bölümü’nde sentezlemiş olduğu metal komplekslerini kullanmamıza imkân tanıyan Prof. Dr. Bülent DEDE’ye teşekkürlerimi sunarım.

Effects of Polynuclear Metal Complexes on Plasmid DNA, Alkaline Protease, and Their Antioxidant Capacities

Abstract

In this work, the influence of previously synthesized compounds on alkaline protease isolated from Bacillus subtilis, their interaction with plasmid DNA, and their antioxidant potential were examined.The compounds evaluated included N′-Bis[1-(4-phenylphenyl)-2-hydroxyimino-2-(pyrrolidino)-1-ethylidene]-1,3-propanediamine along with its heterodinuclear Cu(II)-Mn(II) and Cu(II)-Co(II) complexes, as well as N,N′-Bis[1-(4-phenylphenyl)-2-hydroxyimino-2-(4-chloroanilino)-1-ethylidene]-1,3-propanediamine and its homodinuclear and homotrinuclear Cu(II) complexes. Under optimized production conditions, the enzyme underwent purification utilizing a sequential process of ammonium sulfate precipitation followed by gel filtration chromatography. This process led to a 69.6-fold increase in purity, with an overall recovery yield of 9.9%. SDS-PAGE analysis revealed a single protein band with a molecular weight of approximately 30.0 kD. Metal complexes have been demonstrated to enhance protease activity. Among the complexes under investigation, the homodinuclear Cu(II) complex B2 demonstrated the highest antioxidant activity. Furthermore, the presence of Cu(II) complexes resulted in an augmentation of linear plasmid DNA, an effect that was considerably amplified in the presence of H₂O₂. It was observed that certain complexes were capable of completely degrading DNA, while others were able to disrupt the supercoiled structure upon the addition of hydrogen peroxide.

Keywords

• Alkaline protease
• Bacillus subtilis
• Schiff base
• Antioxidant
• Plasmid DNA.

Project Number

2146-D-10

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