Saccharomyces cerevisiae L. Hücre Kültürü Oksidatif Stres Modelinde Bazı Fosfazen Türevlerinin Biyokimyasal Aktiviteleri
Öz
Bu çalışmada, Saccharomyces cerevisiae L. kültür ortamında bazı fosfazenlerin biyokimyasal aktiviteleri belirlendi. Deneysel uygulamada farklı fosfazen molekülleri kullanıldı. Çalışma kapsamındaki deney grupları; kontrol grubu, H2O2 (Hidrojen peroksit) ve fosfazen molekülleri grupları şeklinde düzenlendi. Gruplar hazırlandıktan sonra, kontrol grubu dışında diğer kültürlere 30 µg fosfazen ve 100 µl H2O2 ilave edildi. 30 °C de 72 saat inkübasyona bırakıldı. İnkübasyon sonunda hücre peletleri ayrıldı. Elde edilen süpernatantdan Glutatyon S-Transferaz (GST) ile total protein düzeyleri belirlendi. Hekzan/izopropanol alkol karışımı ile elde edilen homojenattan da yağ asidi ve lipofilik moleküllerin analizi yapıldı. Deney sonuçlarına göre, fosfazen molekülü ve H2O2 ilave edilen maya hücrelerinde total protein değerleri ile GST değerlerinde paralel bir artış gözlenirken, bazı gruplarda protein miktarında artış saptandığı halde GST düzeyinde azalma olduğu belirlendi. S. cerevisiae’ nin membran yapısında önemli bir yer kaplayan ergosterolün, T3 ve T3B kodlu fosfazenler ile H2O2 gruplarında kontrol grubuna göre yüksek, T4 kodlu fosfazenler ile H2O2 gruplarında ise düşük olduğu belirlendi. Sonuç olarak; S. cerevisiae kültür ortamına fosfazen ve H2O2 moleküllerin eklenmesinin, ergosterol ve yağ asidi sentezi ile yağ asitlerinin hidrokarbon zincirine çift bağ girişi yapan enzimlerin son ürünlerinde artışlara ya da azalışlara neden olduğu ortaya konulmuştur.
Anahtar Kelimeler
Fosfazen H2O2 (Hidrojen peroksit) Lipofilik moleküller Saccharomyces cerevisiae L. Yağ asiti
Destekleyen Kurum
Bitlis Eren Üniversitesi Rektörlüğü Bilimsel Araştırma Projeleri (BAP) birimi
Proje Numarası
BEBAP 2019.008
Teşekkür
Bu çalışma. Bitlis Eren Üniversitesi Rektörlüğü Bilimsel Araştırma Projeleri (BAP) birimi tarafından BEBAP 2019.008 nolu proje numarası ile desteklenmiştir.
Kaynakça
- Allcock, HR.. Morrissey, W.K. & Winograd N. (1996). Controlled Formation of Carboxylic Acid Groups at Polyphosphazene Surfaces; Oxidative and Hydrolytic Routes.Chemistry of Materials. 8, 2730-2738.
- Bergman, L.W. (2001). Growth and Maintenance of Yeast. 2001. Methods in Molecular Biology. Vol. 177. Two Hybrid Systems: Methods and Protocols Edited by: P. N. MacDonald © Humana Press Inc.. Totowa. NJ
- Binici, A.. Okumuş, A.. Yakut, M., Elmas, G., Kılıç, Z., Koyunoğlu, D., Açık, L. & Şimşek, H. (2022). Phosphorus-nitrogen compounds. Part 56. Comparative syntheses and spectral properties of multiheterocyclic 2-cis-4-ansa and spiro-ferrocenyl (N/O)cyclotetraphosphazenes: Antituberculosis and antimicrobial activity and DNA interaction studies. Phosphorus, Sulfur, and Silicon and the Related Elements. 197(1), 18-29.
- Carriedo, GA., Alonso, FJG., Elipe, PG., Brillans, E. & Julia, L. (1996). Incorparation of Stable Organic Radicals into Cyclotriphosphazene: Preparation and Charecterization of Mono – and Diradical Adducts. Journal of the Organic Letters. 3, 1625-1628.
- Christie, WW. (1992). Gas Chromatography and Lipids. Glaskow. The Oil Press.
- Civan, M. (2014). Bazı Trimerik Fosfazen Türevlerinin Sentezi. Kristalografik. Spektroskopik Yöntemler ile Incelenmesi ve Langmuir-Blodgett Tekniği Kullanılarak Ultra-İnce Filmlerinin Hazırlanması. Doctor of Philosophy. Department of Physics Engineering. 358.
- Duan, L.L., Shi, Y., Jiang, R., Yang, Q., Wang, Y.Q., Liu, P.T., Duan, C.Q. & Yan, G.L. (2015). Effects of Adding Unsaturated Fatty Acids on Fatty Acid Composition of Saccharomyces cerevisiae and Major Volatile Compounds in Wine. Centre for Viticulture and Oenology. College of Food Science and Nutritional Engineering. China Agricultural University. Beijing 100083. 36(2), 285-295.
- Dumanoğulları, F. (2006). Bs (Fenoks) Fosfazen Türevlerinin Sentez ve Yapıları. Yüksek Lisans Tezi. Ankara Üniversitesi. Fen Bilimleri Enstitüsü. Kimya Anabilim Dalı. Ankara.
- Erdener Çıralı, D., Dayan, O., Özdemir, N. & Hacıoğlu, N. (2015). A New Phosphazene Derivative, spiro-N3P3[(O2C12H8)2(OC6H6N-3)2], and its Ru(II) complex: Syntheses, crystal structure, catalytic activity and antimicrobial activity studies. Polyhedron. 88, 170-175.
- Greish, Y.E., Bender, J.D., Lakshmi. S., Brown, P.W., Allcock, H.R. & Laurencin, C.T. (2005). Low temperature formation of hydroxyapatite-poly(alkyloxybenzoate)phosphazene composites for biomedical applications. Biomaterials. 26:1–9.
- Habig, W.H., Pabst, M.J. & Jakoby, W.B. (1974). Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. Journal of Biological Chemistry 249(22),7130-7139.
- Hara, A. & Radin, N.S. (1978). Lipid extraction of tissues with a low-toxicity solvent. Analytical Biochemistry. 90(1), 420–426.
- Işıklan, M., Asmafiliz, N., Özalp, E.E., İlter, E.E., Kılıç, Z., Çoşut, B., Yeşilot, S., Kılıç, A., Öztürk., A., Hokelek, T., Koc, L.Y., Acık, L. & Akyüz, E. (2010). Syntheses, Structure İnvestigations, Biological Activities and DNA İnteractions of New N/O Spirocyclic Derivatives. The NMR Behaviors of Chiral Phosphazenes with Stereogenic Centers Upon The Addition of Chiral Solvating Agents. Inorganic Chemistry. 49:7057–7071.
- Kılıç, A., Beğeç, S., Çetinkaya, B., Kılıç, Z., Gündüz, N., Yıldız, M. & Hökelek, T. (1996). Unusual products in the reaction of hexachlorocyclotriphosphazatriene with sodium aryl oxides. Journal of Heteroatom Chemistry. 44(7), 249-256.
- Koçak, S.B., Koçoğlu, S., Okumuş, A., Kılıç, Z., Öztürk, A., Hökelek, T., Öner, Y., & Açık, L. (2013). Syntheses, Spectroscopic Properties, Crystal Structures, Biological Activities and DNA Interactions of Heterocyclic Amine Substituted Spiro-Ansa-Spiro- and Spiro-Bino-Spiro-Phosphazenes. Inorganica Chimica Acta. 406, 160-170.
- Laguna, M.T.R., Tarazona, M.P., Carriedo, G.A., Allonso, F.J.G., Fidalgo, J.I., & Saiz, E. (2002). Thermal degradation and solution properties of poly (2. 2’ – dioxybiphenylphosphazene). Journal of Macromolecules. 35, 7505-7515.
- Lowry, O.H., Rosenbrough, N.J., Farr, A.L., & Randall, R.J. (1951). Protein Measurement with The Folin-Phenol Reagent. The Journal of Biochemistry. 193, 265- 277.
- Okutan, E., Aydın, G.O., Hacıvelioğlu, F., Kılıç, A., Beyaz, S.K., & Yeşilot, S. (2011). Synthesis and Characterization of Soluble Multi-Walled Carbon Nanotube/Poly(Organophosphazene) Composites. Polymer. 52,1241–1248.
- Onder, A., & Ozay, H. (2021). Synthesis and Characterization of Biodegradable and Antioxidant Phpsphazene-Tannic Acid Nanospheres and Their Utilization as Drug carrier Material. Materials Science and Engineering:C. 120, 111723.
- Ozay, H., & Ozay, O. (2014). Synthesis and Characterization of Drug Microspheres Containing Phosphazene for Biomedical Applications. Colloids and Surfaces A: Physicochemical and Engineering Aspects. 450, 99-105.
- Özşahin, A.D., & Bozhan, N. (2018). Bazı Schiff Bazlarının S. cerevisiae BY4741 Kültür Ortamlarında Biyokimyasal Parametreler Üzerine Etkileri. KSÜ Tarım ve Doğa Dergisi. 21(2), 131-140.
- Özşahin, AD., Beşer, D., Öztürk, A.İ., Aslan, F., & Yılmaz, Ö. (2020). Siklofosfazen Türevlerinin Maya Kültür Ortamlarında Malondialdehit, Glutatyon ve Total Protein Miktarları Üzerine Etkileri. KSÜ Tarım ve Doğa Dergisi. 23(2), 281-288.
- Özşahin, A.D., Erdoğdu, A., Kireçci, O.A., & Yılmaz, Ö. (2022). Aromatik Halkalı Bazı Fosfazenlerin S. cerevisiae L. Kültüründe Biyokimyasal Aktivitelerinin Belirlenmesi. KSÜ Tarım ve Doğa Dergisi. 25(2), 234-242.
- Öztürk, A.İ., Yilmaz, O., Kirbag, S., & Arslan, M. (2000). Antimicrobial and Biological Effects of Ipemphos and Amphos on Bacterial and Yeast Strains. Cell Biochemistry and Function. 18(2), 117-126.
- Sazhin, SV., Harrup, M.K., & Gering, K.L. (2011). Characterization of Low-Flammability Electrolytes for Lithium-Ion Batteries. Journal of Power Sources. 196: 3433–3438.
- Siwy, M., Sek, D., Kaczmarczyk, B., Jaroszewicz, I., Nasulewicz, A., Pelczynska, M., Nevozhay, D., & Opolski, A. (2006). Synthesis and In Vitro Antileukemic Activity of Some New 1.3-(Oxytetraethylenoxy) Cyclotriphosphazene Derivatives. Journal of Medicinal Chemistry. 49, 806-810.
- Spencer, J., Phister, T.G., Smart, K.A., & Greetham, D. (2014). Tolerance of Pentose Utilising Yeast to Hydrogen Peroxide-Induced Oxidative Stress. BMC Research Notes. 7, 151.
- Sun, L., Liu, T., Li, H., Yang, L., Meng, L., Lu, Q., & Long, J. (2015). Fluorescent and Cross-Linked Organic−Inorganic Hybrid Nanoshells for Monitoring Drug Delivery. ACS Applied Materials & Interfaces. 7, 4990−4997.
- Şenkuytu, E., Akbaş, N., Yıldırım, T., & Yenilmez Çiftçi, G. (2022). The bioactive new type paraben decorated dispiro-cyclotriphosphaze compounds: synthesis, characterization and cytotoxic activity studies. The Journal of Molecular Structure. 1255, 132438.
- Tvrzicka, E., Vecka, M., Stankova, B., & Zak, A. (2002). Analysis of Fatty Acids Inplasma Lipoproteins by Gas Chromatography Flame Ionisation Detection Quantative Aspects. Analytica Chimica Acta. 465, 337-350
- Uslu, A., & Yeşilot, S. (2015). Chiral configurations in cyclophosphazene chemistry. Coordination Chemistry Reviews. 291, 28-67.
- Xia, Y., Shen, J., Alamri H. Zhan, J., Wang, Y., Zhang, G. (2017). Revealing the Cytotoxicity of Residues of Phosphazene Catalysts Used for the Synthesis of Poly(Ethylene Oxide). Biomacromolecules. 18(10), 3233-3237.
Antioxidant Effects of Some Phosphazene Derivatives in Saccharomyces cerevisiae L. Cell Culture Oxidative Stress Model
Öz
In this study, biochemical activities of some phosphazenes were determined in Saccharomyces cerevisiae L. culture medium. Different phosphazene molecules were used in experimental practice. The experimental groups within the scope of the study were organized as control group, H2O2 (Hydrogen peroxide) and groups of phosphazen molecules. After the groups were prepared, 30 µg of phosphazen and 100 µl of H2O2 were added to other cultures, except for the control group of S. cerevisiae culture. It was incubated for 72 hours at 30 °C. At the end of incubation, cell pellets were separated. Glutathione S-Transferase (GST) and Total protein levels were determined from supernatant. The fatty acid and lipophilic molecules were analyzed from the homogenate obtained with the hexane / isopropanol alcohol mixture. According to our experimental results, while total protein values and GST values increased in parallel with the phosphazen molecule and H2O2 added yeast cells, GST levels were decreased in some groups, although an increase in the amount of protein was observed. Ergosterol, which has an important place in the membrane structure of S. cerevisiae, was found to be higher in T3 and T3B coded phosphazenes and H2O2 groups compared to the control group, and low in T4 coded phosphazenes and H2O2 groups. Our study results revealed that as a result of the addition of phosphazen and H2O2 molecules to the culture medium of S. cerevisiae, ergosterol and fatty acid synthesis, fatty acids cause increases or decreases in the end products of enzymes that double-enter the hydrocarbon chain.
Anahtar Kelimeler
Phosphazene H2O2 (hydrogen peroxide) Lipophilic molecules Saccharomyces cerevisiae L. Fatty acid
Proje Numarası
BEBAP 2019.008
Kaynakça
- Allcock, HR.. Morrissey, W.K. & Winograd N. (1996). Controlled Formation of Carboxylic Acid Groups at Polyphosphazene Surfaces; Oxidative and Hydrolytic Routes.Chemistry of Materials. 8, 2730-2738.
- Bergman, L.W. (2001). Growth and Maintenance of Yeast. 2001. Methods in Molecular Biology. Vol. 177. Two Hybrid Systems: Methods and Protocols Edited by: P. N. MacDonald © Humana Press Inc.. Totowa. NJ
- Binici, A.. Okumuş, A.. Yakut, M., Elmas, G., Kılıç, Z., Koyunoğlu, D., Açık, L. & Şimşek, H. (2022). Phosphorus-nitrogen compounds. Part 56. Comparative syntheses and spectral properties of multiheterocyclic 2-cis-4-ansa and spiro-ferrocenyl (N/O)cyclotetraphosphazenes: Antituberculosis and antimicrobial activity and DNA interaction studies. Phosphorus, Sulfur, and Silicon and the Related Elements. 197(1), 18-29.
- Carriedo, GA., Alonso, FJG., Elipe, PG., Brillans, E. & Julia, L. (1996). Incorparation of Stable Organic Radicals into Cyclotriphosphazene: Preparation and Charecterization of Mono – and Diradical Adducts. Journal of the Organic Letters. 3, 1625-1628.
- Christie, WW. (1992). Gas Chromatography and Lipids. Glaskow. The Oil Press.
- Civan, M. (2014). Bazı Trimerik Fosfazen Türevlerinin Sentezi. Kristalografik. Spektroskopik Yöntemler ile Incelenmesi ve Langmuir-Blodgett Tekniği Kullanılarak Ultra-İnce Filmlerinin Hazırlanması. Doctor of Philosophy. Department of Physics Engineering. 358.
- Duan, L.L., Shi, Y., Jiang, R., Yang, Q., Wang, Y.Q., Liu, P.T., Duan, C.Q. & Yan, G.L. (2015). Effects of Adding Unsaturated Fatty Acids on Fatty Acid Composition of Saccharomyces cerevisiae and Major Volatile Compounds in Wine. Centre for Viticulture and Oenology. College of Food Science and Nutritional Engineering. China Agricultural University. Beijing 100083. 36(2), 285-295.
- Dumanoğulları, F. (2006). Bs (Fenoks) Fosfazen Türevlerinin Sentez ve Yapıları. Yüksek Lisans Tezi. Ankara Üniversitesi. Fen Bilimleri Enstitüsü. Kimya Anabilim Dalı. Ankara.
- Erdener Çıralı, D., Dayan, O., Özdemir, N. & Hacıoğlu, N. (2015). A New Phosphazene Derivative, spiro-N3P3[(O2C12H8)2(OC6H6N-3)2], and its Ru(II) complex: Syntheses, crystal structure, catalytic activity and antimicrobial activity studies. Polyhedron. 88, 170-175.
- Greish, Y.E., Bender, J.D., Lakshmi. S., Brown, P.W., Allcock, H.R. & Laurencin, C.T. (2005). Low temperature formation of hydroxyapatite-poly(alkyloxybenzoate)phosphazene composites for biomedical applications. Biomaterials. 26:1–9.
- Habig, W.H., Pabst, M.J. & Jakoby, W.B. (1974). Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. Journal of Biological Chemistry 249(22),7130-7139.
- Hara, A. & Radin, N.S. (1978). Lipid extraction of tissues with a low-toxicity solvent. Analytical Biochemistry. 90(1), 420–426.
- Işıklan, M., Asmafiliz, N., Özalp, E.E., İlter, E.E., Kılıç, Z., Çoşut, B., Yeşilot, S., Kılıç, A., Öztürk., A., Hokelek, T., Koc, L.Y., Acık, L. & Akyüz, E. (2010). Syntheses, Structure İnvestigations, Biological Activities and DNA İnteractions of New N/O Spirocyclic Derivatives. The NMR Behaviors of Chiral Phosphazenes with Stereogenic Centers Upon The Addition of Chiral Solvating Agents. Inorganic Chemistry. 49:7057–7071.
- Kılıç, A., Beğeç, S., Çetinkaya, B., Kılıç, Z., Gündüz, N., Yıldız, M. & Hökelek, T. (1996). Unusual products in the reaction of hexachlorocyclotriphosphazatriene with sodium aryl oxides. Journal of Heteroatom Chemistry. 44(7), 249-256.
- Koçak, S.B., Koçoğlu, S., Okumuş, A., Kılıç, Z., Öztürk, A., Hökelek, T., Öner, Y., & Açık, L. (2013). Syntheses, Spectroscopic Properties, Crystal Structures, Biological Activities and DNA Interactions of Heterocyclic Amine Substituted Spiro-Ansa-Spiro- and Spiro-Bino-Spiro-Phosphazenes. Inorganica Chimica Acta. 406, 160-170.
- Laguna, M.T.R., Tarazona, M.P., Carriedo, G.A., Allonso, F.J.G., Fidalgo, J.I., & Saiz, E. (2002). Thermal degradation and solution properties of poly (2. 2’ – dioxybiphenylphosphazene). Journal of Macromolecules. 35, 7505-7515.
- Lowry, O.H., Rosenbrough, N.J., Farr, A.L., & Randall, R.J. (1951). Protein Measurement with The Folin-Phenol Reagent. The Journal of Biochemistry. 193, 265- 277.
- Okutan, E., Aydın, G.O., Hacıvelioğlu, F., Kılıç, A., Beyaz, S.K., & Yeşilot, S. (2011). Synthesis and Characterization of Soluble Multi-Walled Carbon Nanotube/Poly(Organophosphazene) Composites. Polymer. 52,1241–1248.
- Onder, A., & Ozay, H. (2021). Synthesis and Characterization of Biodegradable and Antioxidant Phpsphazene-Tannic Acid Nanospheres and Their Utilization as Drug carrier Material. Materials Science and Engineering:C. 120, 111723.
- Ozay, H., & Ozay, O. (2014). Synthesis and Characterization of Drug Microspheres Containing Phosphazene for Biomedical Applications. Colloids and Surfaces A: Physicochemical and Engineering Aspects. 450, 99-105.
- Özşahin, A.D., & Bozhan, N. (2018). Bazı Schiff Bazlarının S. cerevisiae BY4741 Kültür Ortamlarında Biyokimyasal Parametreler Üzerine Etkileri. KSÜ Tarım ve Doğa Dergisi. 21(2), 131-140.
- Özşahin, AD., Beşer, D., Öztürk, A.İ., Aslan, F., & Yılmaz, Ö. (2020). Siklofosfazen Türevlerinin Maya Kültür Ortamlarında Malondialdehit, Glutatyon ve Total Protein Miktarları Üzerine Etkileri. KSÜ Tarım ve Doğa Dergisi. 23(2), 281-288.
- Özşahin, A.D., Erdoğdu, A., Kireçci, O.A., & Yılmaz, Ö. (2022). Aromatik Halkalı Bazı Fosfazenlerin S. cerevisiae L. Kültüründe Biyokimyasal Aktivitelerinin Belirlenmesi. KSÜ Tarım ve Doğa Dergisi. 25(2), 234-242.
- Öztürk, A.İ., Yilmaz, O., Kirbag, S., & Arslan, M. (2000). Antimicrobial and Biological Effects of Ipemphos and Amphos on Bacterial and Yeast Strains. Cell Biochemistry and Function. 18(2), 117-126.
- Sazhin, SV., Harrup, M.K., & Gering, K.L. (2011). Characterization of Low-Flammability Electrolytes for Lithium-Ion Batteries. Journal of Power Sources. 196: 3433–3438.
- Siwy, M., Sek, D., Kaczmarczyk, B., Jaroszewicz, I., Nasulewicz, A., Pelczynska, M., Nevozhay, D., & Opolski, A. (2006). Synthesis and In Vitro Antileukemic Activity of Some New 1.3-(Oxytetraethylenoxy) Cyclotriphosphazene Derivatives. Journal of Medicinal Chemistry. 49, 806-810.
- Spencer, J., Phister, T.G., Smart, K.A., & Greetham, D. (2014). Tolerance of Pentose Utilising Yeast to Hydrogen Peroxide-Induced Oxidative Stress. BMC Research Notes. 7, 151.
- Sun, L., Liu, T., Li, H., Yang, L., Meng, L., Lu, Q., & Long, J. (2015). Fluorescent and Cross-Linked Organic−Inorganic Hybrid Nanoshells for Monitoring Drug Delivery. ACS Applied Materials & Interfaces. 7, 4990−4997.
- Şenkuytu, E., Akbaş, N., Yıldırım, T., & Yenilmez Çiftçi, G. (2022). The bioactive new type paraben decorated dispiro-cyclotriphosphaze compounds: synthesis, characterization and cytotoxic activity studies. The Journal of Molecular Structure. 1255, 132438.
- Tvrzicka, E., Vecka, M., Stankova, B., & Zak, A. (2002). Analysis of Fatty Acids Inplasma Lipoproteins by Gas Chromatography Flame Ionisation Detection Quantative Aspects. Analytica Chimica Acta. 465, 337-350
- Uslu, A., & Yeşilot, S. (2015). Chiral configurations in cyclophosphazene chemistry. Coordination Chemistry Reviews. 291, 28-67.
- Xia, Y., Shen, J., Alamri H. Zhan, J., Wang, Y., Zhang, G. (2017). Revealing the Cytotoxicity of Residues of Phosphazene Catalysts Used for the Synthesis of Poly(Ethylene Oxide). Biomacromolecules. 18(10), 3233-3237.
Ayrıntılar
| Birincil Dil | Türkçe |
|---|---|
| Konular | Yapısal Biyoloji |
| Bölüm | ARAŞTIRMA MAKALESİ (Research Article) |
| Yazarlar | |
| Proje Numarası | BEBAP 2019.008 |
| Yayımlanma Tarihi | 28 Şubat 2023 |
| Gönderilme Tarihi | 31 Aralık 2021 |
| Kabul Tarihi | 10 Mart 2022 |
| Yayımlandığı Sayı | Yıl 2023Cilt: 26 Sayı: 1 |
Kaynak Göster
2022-JIF = 0.500
2022-JCI = 0.170
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