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

Yıl 2026, Cilt: 30 Sayı: 1 , 51 - 58 , 24.04.2026
https://doi.org/10.19113/sdufenbed.1734812
https://izlik.org/JA96UD72JW

Öz

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.

Etik Beyan

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.

Destekleyen Kurum

Süleyman Demirel Üniversitesi Bilimsel Araştırma Projeleri Yönetim Birimi

Proje Numarası

2146-D-10

Teşekkür

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.

Kaynakça

  • [1] Adinarayana, K., Ellaiah, P., Prasad, D. S. 2003. Purification and partial characterization of thermostable serine alkaline protease from a newly isolated Bacillus subtilis PE-11. AAPS PharmSciTech, 4(4), 440–448.
  • [2] Patel, R. K., Dodia, M. S., Joshi, R. H., Singh, S. P. 2006. Purification and characterization of alkaline protease from a newly isolated haloalkaliphilic Bacillus sp. Process Biochemistry, 41(9), 2002–2009.
  • [3] Zambare, V. 2010. Purification and Characterization of Neutral Serine Protease from Bacillus sp. Asiatic Journal of Biotechnol. Resources, 3, 183–192.
  • [4] Beg, K. Q., Gupta, R. 2003. Purification and characterization of an oxidation-stable, thiol-dependent serine alkaline protease from Bacillus mojavensis. Enzyme and Microbial Technology, 32(2), 294–304.
  • [5] Singh, J., Vohra, R. M., Sahoo, D. K. 2001. Purification and characterization of two extracellular alkaline proteases from a newly isolated obligate alkalophilic Bacillus sphaericus. Journal of Industrial Microbiology & Biotechnology, 26(6), 387–393.
  • [6] Huang, Q., Peng, Y., Li, X., Wang, H., Zhang, Y. 2003. Purification and characterization of an extracellular alkaline serine protease with dehairing function from Bacillus pumilus. Current Microbiology, 46(3), 169–173.
  • [7] Joo, H. S., Chang, C. S. 2005. Production of protease from a new alkalophilic Bacillus sp. I-312 grown on soybean meal: optimization and some properties. Process Biochemistry, 40(3–4), 1263–1270.
  • [8] Sevinç, N., Demirkan, E. 2011. Production of Protease by Bacillus sp. N-40 Isolated from Soil and Its Enzymatic Properties. Journal of Biology of the Environment Sciences, 5(14), 95–103.
  • [9] Nilegaonkar, S. S., lowry, V. P., Kanekar, P. P., Dhakephalkar, P. K., Sarnaik, S. S., Chopade, B. A. 2007. Production and partial characterization of dehairing protease from Bacillus cereus MCM B-326. Bioresource Technology, 98(7), 1238–1245.
  • [10] Gupta, R., Beg, Q. K., Khan, S., Chauhan, B. 2005. One-step purification and characterization of an alkaline protease from haloalkaliphilic Bacillus sp. Journal of Chromatography A, 1075(1–2), 103–108.
  • [11] Johnvesly, B., Naik, G. R. 2001. Production of thermostable alkaline protease from thermophilic and alkaliphilic Bacillus sp. JB-99 in a chemically defined medium. Process Biochemistry, 37(2), 139–144.
  • [12] Dede, B., Karipcin, F., Cengiz, M. 2009. Novel homo- and hetero-nuclear copper(II) complexes of tetradentate Schiff bases: Synthesis, characterization, solvent-extraction and catalase-like activity studies. Journal of Hazardous Materials, 163, 1148–1156.
  • [13] Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227(5259), 680–685.
  • [14] Anson, M.L. (1938) The Estimation of Pepsin, Trypsin, Papain, and Cathepsin with Hemoglobin. The Journal of General Physiology, 22, 79-89.
  • [15] Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J. 1951. Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry, 193, 265–275.
  • [16] Brand-Williams, W., Cuvelier, M.E., Berset, C., 1995. Use Of A Free Radical Method To Evaluate Antioxidant Activity. LWT - Food Science and Technology. Volume 28(1), 25-30.
  • [17] Fukumoto, L.R., & Mazza, G., 2000. Assessing antioxidant and prooxidant activities of phenolic compounds. Journal of agricultural and food chemistry. 48(8), 3597-3604.
  • [18] Mates J M, Perez-Gomez C & Nunez de Castro I (1999). Antioxidant enzymes and human diseases. Clin. Biochem. 32: 595–603.
  • [19] Mei, C., Jiang, X. 2005. A novel surfactant- and oxidation-stable alkaline protease from Vibrio metschnikovii DL 33-51. Process Biochemistry, 40(6), 2167–2172.
  • [20] Yang, J., Shih, I., Tzeng, Y., Wang, S. 2000. Production and purification of protease from a Bacillus subtilis that can deproteinize crustacean wastes. Enzyme and Microbial Technology, 26(5–6), 406–413.
  • [21] Reddy, K. H., Reddy, P. S. 2000. Nuclease Activity of Mixed Ligand Complexes of Copper (II) with Heteroaromatic Derivatives and Picoline. Transition Metal Chemistry, 25(5), 505–510.
  • [22] Sağlam, N., Çolak, A., Serbest, K., Dülger, S., Güner, S., Karaböcek, S., Beldüz, A. O. 2002. Oxidative Cleavage of DNA by Homo- and Heteronuclear Cu(II)-Mn(II) Complexes of an Oxime-Type Ligand. Biometals, 15(4), 357–365.
  • [23] Ceyhan, G., Çelik, C., Uruş, S., Demirtaş, İ., Elmastaş, M., Tümer, M. 2011. Antioxidant, electrochemical, thermal, antimicrobial and alkane oxidation properties of tridentate Schiff base ligands and their metal complexes. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 81(1), 184–198.
  • [24] Zhang, Y., Zou, B., Chen, Z., Pan, Y., Wang, H., Liang, H., Yi, X. 2011. Synthesis and antioxidant activities of novel 4-Schiff base-7-benzyloxy-coumarin derivatives. Bioorganic & Medicinal Chemistry Letters, 21(22), 6811–6815.
  • [25] Tian, M., Ihmels, H., Brötz, E. 2010. DNA cleavage by the Cu (ii) complex of the DNA-intercalating 9-bis (pyridin-2-ylmethyl) aminobenzo [b] quinolizinium. Dalton Transactions, 39(35), 8195-8202.
  • [26] Massoud, S. S., Louka, F. R., Xu, W., Perkins, R. S., Vicente, R., Albering, J. H., Mautner, F. A. 2011. DNA cleavage by structurally characterized dinuclear copper (II) complexes based on triazine. European Journal of Inorganic Chemistry, 2011(23), 3469–3479.
  • [27] Kim, J. H., Kim, S. H. 2003. Double Strand DNA Cleavage by a Dinuclear Cu(II) Complex. Chemistry Letters, 32(6), 490.

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

Yıl 2026, Cilt: 30 Sayı: 1 , 51 - 58 , 24.04.2026
https://doi.org/10.19113/sdufenbed.1734812
https://izlik.org/JA96UD72JW

Öz

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.

Proje Numarası

2146-D-10

Kaynakça

  • [1] Adinarayana, K., Ellaiah, P., Prasad, D. S. 2003. Purification and partial characterization of thermostable serine alkaline protease from a newly isolated Bacillus subtilis PE-11. AAPS PharmSciTech, 4(4), 440–448.
  • [2] Patel, R. K., Dodia, M. S., Joshi, R. H., Singh, S. P. 2006. Purification and characterization of alkaline protease from a newly isolated haloalkaliphilic Bacillus sp. Process Biochemistry, 41(9), 2002–2009.
  • [3] Zambare, V. 2010. Purification and Characterization of Neutral Serine Protease from Bacillus sp. Asiatic Journal of Biotechnol. Resources, 3, 183–192.
  • [4] Beg, K. Q., Gupta, R. 2003. Purification and characterization of an oxidation-stable, thiol-dependent serine alkaline protease from Bacillus mojavensis. Enzyme and Microbial Technology, 32(2), 294–304.
  • [5] Singh, J., Vohra, R. M., Sahoo, D. K. 2001. Purification and characterization of two extracellular alkaline proteases from a newly isolated obligate alkalophilic Bacillus sphaericus. Journal of Industrial Microbiology & Biotechnology, 26(6), 387–393.
  • [6] Huang, Q., Peng, Y., Li, X., Wang, H., Zhang, Y. 2003. Purification and characterization of an extracellular alkaline serine protease with dehairing function from Bacillus pumilus. Current Microbiology, 46(3), 169–173.
  • [7] Joo, H. S., Chang, C. S. 2005. Production of protease from a new alkalophilic Bacillus sp. I-312 grown on soybean meal: optimization and some properties. Process Biochemistry, 40(3–4), 1263–1270.
  • [8] Sevinç, N., Demirkan, E. 2011. Production of Protease by Bacillus sp. N-40 Isolated from Soil and Its Enzymatic Properties. Journal of Biology of the Environment Sciences, 5(14), 95–103.
  • [9] Nilegaonkar, S. S., lowry, V. P., Kanekar, P. P., Dhakephalkar, P. K., Sarnaik, S. S., Chopade, B. A. 2007. Production and partial characterization of dehairing protease from Bacillus cereus MCM B-326. Bioresource Technology, 98(7), 1238–1245.
  • [10] Gupta, R., Beg, Q. K., Khan, S., Chauhan, B. 2005. One-step purification and characterization of an alkaline protease from haloalkaliphilic Bacillus sp. Journal of Chromatography A, 1075(1–2), 103–108.
  • [11] Johnvesly, B., Naik, G. R. 2001. Production of thermostable alkaline protease from thermophilic and alkaliphilic Bacillus sp. JB-99 in a chemically defined medium. Process Biochemistry, 37(2), 139–144.
  • [12] Dede, B., Karipcin, F., Cengiz, M. 2009. Novel homo- and hetero-nuclear copper(II) complexes of tetradentate Schiff bases: Synthesis, characterization, solvent-extraction and catalase-like activity studies. Journal of Hazardous Materials, 163, 1148–1156.
  • [13] Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227(5259), 680–685.
  • [14] Anson, M.L. (1938) The Estimation of Pepsin, Trypsin, Papain, and Cathepsin with Hemoglobin. The Journal of General Physiology, 22, 79-89.
  • [15] Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J. 1951. Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry, 193, 265–275.
  • [16] Brand-Williams, W., Cuvelier, M.E., Berset, C., 1995. Use Of A Free Radical Method To Evaluate Antioxidant Activity. LWT - Food Science and Technology. Volume 28(1), 25-30.
  • [17] Fukumoto, L.R., & Mazza, G., 2000. Assessing antioxidant and prooxidant activities of phenolic compounds. Journal of agricultural and food chemistry. 48(8), 3597-3604.
  • [18] Mates J M, Perez-Gomez C & Nunez de Castro I (1999). Antioxidant enzymes and human diseases. Clin. Biochem. 32: 595–603.
  • [19] Mei, C., Jiang, X. 2005. A novel surfactant- and oxidation-stable alkaline protease from Vibrio metschnikovii DL 33-51. Process Biochemistry, 40(6), 2167–2172.
  • [20] Yang, J., Shih, I., Tzeng, Y., Wang, S. 2000. Production and purification of protease from a Bacillus subtilis that can deproteinize crustacean wastes. Enzyme and Microbial Technology, 26(5–6), 406–413.
  • [21] Reddy, K. H., Reddy, P. S. 2000. Nuclease Activity of Mixed Ligand Complexes of Copper (II) with Heteroaromatic Derivatives and Picoline. Transition Metal Chemistry, 25(5), 505–510.
  • [22] Sağlam, N., Çolak, A., Serbest, K., Dülger, S., Güner, S., Karaböcek, S., Beldüz, A. O. 2002. Oxidative Cleavage of DNA by Homo- and Heteronuclear Cu(II)-Mn(II) Complexes of an Oxime-Type Ligand. Biometals, 15(4), 357–365.
  • [23] Ceyhan, G., Çelik, C., Uruş, S., Demirtaş, İ., Elmastaş, M., Tümer, M. 2011. Antioxidant, electrochemical, thermal, antimicrobial and alkane oxidation properties of tridentate Schiff base ligands and their metal complexes. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 81(1), 184–198.
  • [24] Zhang, Y., Zou, B., Chen, Z., Pan, Y., Wang, H., Liang, H., Yi, X. 2011. Synthesis and antioxidant activities of novel 4-Schiff base-7-benzyloxy-coumarin derivatives. Bioorganic & Medicinal Chemistry Letters, 21(22), 6811–6815.
  • [25] Tian, M., Ihmels, H., Brötz, E. 2010. DNA cleavage by the Cu (ii) complex of the DNA-intercalating 9-bis (pyridin-2-ylmethyl) aminobenzo [b] quinolizinium. Dalton Transactions, 39(35), 8195-8202.
  • [26] Massoud, S. S., Louka, F. R., Xu, W., Perkins, R. S., Vicente, R., Albering, J. H., Mautner, F. A. 2011. DNA cleavage by structurally characterized dinuclear copper (II) complexes based on triazine. European Journal of Inorganic Chemistry, 2011(23), 3469–3479.
  • [27] Kim, J. H., Kim, S. H. 2003. Double Strand DNA Cleavage by a Dinuclear Cu(II) Complex. Chemistry Letters, 32(6), 490.
Toplam 27 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Konular Enzimler, Proteinler ve Peptitler
Bölüm Araştırma Makalesi
Yazarlar

Şevkinaz Doğan 0000-0001-6180-7586

İsmail Özmen 0000-0002-5457-8188

Proje Numarası 2146-D-10
Gönderilme Tarihi 4 Temmuz 2025
Kabul Tarihi 9 Mart 2026
Yayımlanma Tarihi 24 Nisan 2026
DOI https://doi.org/10.19113/sdufenbed.1734812
IZ https://izlik.org/JA96UD72JW
Yayımlandığı Sayı Yıl 2026 Cilt: 30 Sayı: 1

Kaynak Göster

APA Doğan, Ş., & Özmen, İ. (2026). Polinükleer Metal Komplekslerin Plazmid DNA, Alkalen Proteaz Üzerine Etkileri ve Antioksidan Kapasiteleri. Süleyman Demirel Üniversitesi Fen Bilimleri Enstitüsü Dergisi, 30(1), 51-58. https://doi.org/10.19113/sdufenbed.1734812
AMA 1.Doğan Ş, Özmen İ. Polinükleer Metal Komplekslerin Plazmid DNA, Alkalen Proteaz Üzerine Etkileri ve Antioksidan Kapasiteleri. Süleyman Demirel Üniv. Fen Bilim. Enst. Derg. 2026;30(1):51-58. doi:10.19113/sdufenbed.1734812
Chicago Doğan, Şevkinaz, ve İsmail Özmen. 2026. “Polinükleer Metal Komplekslerin Plazmid DNA, Alkalen Proteaz Üzerine Etkileri ve Antioksidan Kapasiteleri”. Süleyman Demirel Üniversitesi Fen Bilimleri Enstitüsü Dergisi 30 (1): 51-58. https://doi.org/10.19113/sdufenbed.1734812.
EndNote Doğan Ş, Özmen İ (01 Nisan 2026) Polinükleer Metal Komplekslerin Plazmid DNA, Alkalen Proteaz Üzerine Etkileri ve Antioksidan Kapasiteleri. Süleyman Demirel Üniversitesi Fen Bilimleri Enstitüsü Dergisi 30 1 51–58.
IEEE [1]Ş. Doğan ve İ. Özmen, “Polinükleer Metal Komplekslerin Plazmid DNA, Alkalen Proteaz Üzerine Etkileri ve Antioksidan Kapasiteleri”, Süleyman Demirel Üniv. Fen Bilim. Enst. Derg., c. 30, sy 1, ss. 51–58, Nis. 2026, doi: 10.19113/sdufenbed.1734812.
ISNAD Doğan, Şevkinaz - Özmen, İsmail. “Polinükleer Metal Komplekslerin Plazmid DNA, Alkalen Proteaz Üzerine Etkileri ve Antioksidan Kapasiteleri”. Süleyman Demirel Üniversitesi Fen Bilimleri Enstitüsü Dergisi 30/1 (01 Nisan 2026): 51-58. https://doi.org/10.19113/sdufenbed.1734812.
JAMA 1.Doğan Ş, Özmen İ. Polinükleer Metal Komplekslerin Plazmid DNA, Alkalen Proteaz Üzerine Etkileri ve Antioksidan Kapasiteleri. Süleyman Demirel Üniv. Fen Bilim. Enst. Derg. 2026;30:51–58.
MLA Doğan, Şevkinaz, ve İsmail Özmen. “Polinükleer Metal Komplekslerin Plazmid DNA, Alkalen Proteaz Üzerine Etkileri ve Antioksidan Kapasiteleri”. Süleyman Demirel Üniversitesi Fen Bilimleri Enstitüsü Dergisi, c. 30, sy 1, Nisan 2026, ss. 51-58, doi:10.19113/sdufenbed.1734812.
Vancouver 1.Şevkinaz Doğan, İsmail Özmen. Polinükleer Metal Komplekslerin Plazmid DNA, Alkalen Proteaz Üzerine Etkileri ve Antioksidan Kapasiteleri. Süleyman Demirel Üniv. Fen Bilim. Enst. Derg. 01 Nisan 2026;30(1):51-8. doi:10.19113/sdufenbed.1734812

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