Araştırma Makalesi
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Purification, characterization and dertermination of kinetic features of carbonic anhydrase from turbot (Psetta Maxima) muscle tissue

Yıl 2023, Cilt: 1 Sayı: 1, 1 - 7, 18.06.2023

Öz

In this study, firstly, carbonic anhydrase purification from turbot (Psetta maxima) muscle tissue, together with analysis of the kinetic behaviour and some enzyme properties are described. The purification steps comprised hemolssate preparation, Sepharose-4B-L tyrosine-sulfanilamide affinity gel chromatography and dialyze. The yield was 69.05% and the enzyme was found to be specific activity of 755.2 EU/mg proteins. The overall purification was about 50.65-fold. Temperature of +40C was maintained during the purification process. The molecular mass of the subunit was determined to be 29.7 kDa by SDS polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme had an optimal pH at 8.0 and stable pH at 8.0 and optimal temperature at 30ºC. Km and Vmax for p-nitrophenylacetate as a substrate were also determined.

Kaynakça

  • Badger, M.R., & Price, G.D. (1994). The role of carbonic anhydrase in photosynthesis. Annual Review of Plant Physiology and Plant Molecular Biology, 45, 369-392. https://doi.org/10.1146/annurev.pp.45.060194.002101
  • Bayram, E., Senturk, M., Kufrevioglu, O.I., & Supuran, C.T. (2008). In vitro inhibition of salicylic acid derivatives on human cytosolic carbonic anhydrase isozymes I and II. Bioorganic & Medicinal Chemistry, 16(20), 9101-9105. https://doi.org/10.1016/j.bmc.2008.09.028
  • Bradford, M.M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248. https://doi.org/10.1016/0003-2697(76)90527-3
  • Ceyhun, S.B., Senturk, M., Yerlikaya, E., Erdogan, O., Kufrevioglu, O.I., & Ekinci, D. (2011). Purification and characterization of carbonic anhydrase from the teleost fish Dicentrarchus labrax (European seabass) liver and toxicological effects of metals on enzyme activity. Environmental Toxicology and Pharmacology, 32(1), 69-74. https://doi.org/10.1016/j.etap.2011.03.013
  • Chegwidden, W.R., Edwards, Y., & Carter, N. (2000). The Carbonic Anhydrase-New Horizons. Molecular Bases of Inherited Disease (Scriver, C.R., Beaudet, A. L., Sly, W. S., and Valle, D., eds) 8th Ed., pp. 2165-2204, McGraw-Hill, Inc., Newyork.
  • Clare, B.W., & Supuran, C.T. (2000). Carbonic Anhydrase Inhibitors. Part 86. A QSAR study on some sulfonamide drugs which lower intra-ocular pressure, using the ACE non-linear statistical method. The European Journal of Medicinal Chemistry, 35, 859-865. https://doi.org/10.1016/S0223-5234(00)00182-3
  • Del Prete, S., Vullo, D., De Luca, V., Carginale, V., Osman, S.M., AlOthman, Z., Supuran, C.T., & Capasso, C. (2016). Cloning, expression, purification and sulfonamide inhibition profile of the complete domain of the η-carbonic anhydrase from Plasmodium falciparum. Bioorganic & Medicinal Chemistry Letters, 26(17), 4184-90. https://doi.org/10.1016/j.bmcl.2016.07.060
  • Demirdağ, R., Çomaklı, V., Şentürk, M., Ekinci, D., İrfan Küfrevioğlu, Ö., & Supuran, C.T. (2013). Purification and characterization of carbonic anhydrase from sheep kidney and effects of sulfonamides on enzyme activity. Bioorganic & medicinal chemistry, 21(6), 1522-1525. https://doi.org/10.1016/j.bmc.2012.08.018
  • Ekinci, D., Cavdar, H., Talaz, O., Senturk, M., & Supuran, C.T. (2010). NO-releasing esters show carbonic anhydrase inhibitory action against human isoforms I and II. Bioorganic & Medicinal Chemistry, 18(10), 3559-3563. https://doi.org/10.1016/j.bmc.2010.03.082
  • FAO (2010). The State of World Fisheries and Aquaculture. http://www.fao.org/docrep/013/i1820e/i1820e.pdf Göçer, H., Akıncıoğlu, A., Göksu, S., Gülcin, I., & Supuran, C.T. (2015). Carbonic anhydrase and acetylcholine esterase inhibitory effects of carbamates andsulfamoylcarbamates. Journal of enzyme inhibition and medicinal chemistry, 30(2), 316-320. https://doi.org/10.3109/14756366.2014.928704
  • Göksu, S., Naderi, A., Akbaba, Y., Kalın, P., Akıncıoglu, A., Gulcin, I., Durdagi, S., & Salmas, R.E. (2014). Carbonic anhydrase inhibitory properties of novelbenzylsulfamides using molecular modeling and experimental studies. Bioorganic chemistry, 56, 75-82. https://doi.org/10.1016/j.bioorg.2014.07.009
  • Graham, D., Reed, M.L., Patterson, B.D., Hockley, D.G., & Dwyer, M. (1984). Chemical properties, distribution and physiology of plant and algal carbonic anhydrases. Annals of the New York Academy of Sciences, 429, 222-237. https://doi.org/10.1111/j.1749-6632.1984.tb12340.x
  • Hara, S. (2001). Karadeniz kalkan balıgından (Psetta maxima) döllenmis yumurta ve larva elde edilmesi. SÜMAE, Yunus Arastırma Bülteni, 1,1.
  • Henry, R.P., Smatresk, N.J., & Cameron, J.N. (1988). The distribition of brachial carbonic anhydrase and the effect of gill and erythrocyte carbonic anhydrase inhibition in the cannel catfish, Ictalurus punctatus. Journal of Experimental Biology, 134, 201-218. https://doi.org/10.1242/jeb.134.1.201
  • Jo, B.H., & Hwang, I.S. (2019). Characterization and High-Level Periplasmic Expression of Thermostable α-Carbonic Anhydrase from Thermosulfurimonas Dismutans in Escherichia Coli for CO2 Capture and Utilization. International Journal of Molecular Sciences, 21(1), 103. https://doi.org/10.3390/ijms21010103
  • Jones, A. (1974). Sexual Maturity, Fecundity and Growth of the Turbot (Scophthalmus maximus L.). Journal of the Marine Biological Association of the United Kingdom, 54, 109-125. https://doi.org/10.1017/S0025315400022104
  • Kaya, E.D., Söyüt, H., & Beydemir, S. (2013). Carbonic anhydrase activity from thegilthead sea bream (Sparus aurata) liver: the toxicological effects of heavymetals. Environmental Toxicology and Pharmacology, 36, 514-521. https://doi.org/10.1016/j.etap.2013.05.019
  • Kucuk, M., & Gulcin, İ. (2016). Purification and characterization of the carbonic anhydrase enzyme from Black Sea trout (Salmo trutta Labrax Coruhensis) kidney and inhibition effects of some metal ions on enzyme activity. Environmental Toxicology and Pharmacology, 44, 134-9. https://doi.org/10.1016/j.etap.2016.04.011
  • Laemmli, D.K. (1970). Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature, 227, 680.
  • Maheshwari, N., Kumar, M., Thakur, I.S., & Srivastava, S. (2019). Cloning, expression and characterization of β- and γ‑carbonic anhydrase from Bacillus sp. SS105 for biomimetic sequestration of CO2. International journal of biological macromolecules, 131, 445-452. https://doi.org/10.1016/j.ijbiomac.2019.03.082
  • Memis, D. (2010). Deniz Balıkları Yetistiriciligi. Filiz Kitabevi, İstanbul, 152-172.
  • Munroe, T.A. (2005). Systematic Diversity of the Pleuronectiformes in: Flatfishes
  • Öztürk Sarikaya, S.B., Topal, F., Şentürk, M., Gülçin, İ., & Supuran, C.T. (2011). In vitro inhibition of a-carbonic anhydrase isozymes by some phenolic compounds. Bioorganic & Medicinal Chemistry Letters, 21, 4259. https://doi.org/10.1016/j.bmcl.2011.05.071
  • Peterson, R.E., Tu, C., & Linser, P.J. (1997). Isolation and characterisation of a carbonicanhyrdase homologue from the zebrafish (Danio rerio). Journal of Molecular Evolution, 448, 432-439. https://doi.org/10.1007/pl00006163
  • Rizzello, A., Ciardiello, M.A., Acierno, R., Carratore, V., Verri, T., Di Prisco, G., Storelli, C., & Maffia, M. (2007). Biochemical characterization of a s-glutathionylatedcarbonic anhydrase isolated from gills of the antarctic icefish. The Protein Journal, 26, 335-348. https://doi.org/10.1007/s10930-007-9076-1
  • Samsun, N. (2004). Sinop yöresinde avlanan kalkan (Scophthalmus maeoticus Pallas,1811) balıklarının bazı biyolojik özelliklerinin belirlenmesi üzerine bir arastırma. Ondokuz Mayıs Üniversitesi Fen Bilimleri Enstitüsü Su Ürünleri Avlama ve İşleme Teknolojisi Anabilim Dalı, Doktora Tezi, 171s., Samsun.
  • Segel, I.H. (1968). Biochemical calculations, second edition, Publised by, John Wiley and Sons Inc. New York, USA, 458 pp., ISBN: 0-471-77421-9
  • Sevgili, H., & Nezaki, G. (2010). Turbot Culture in France and Spain. Special Publication 5. February Emre Y. (Eds).
  • Solis, C., Olivera, A., Andrade, E., Ruvalcaba-Sil, J.L., Romero, I., & Celis, H. (1999). PIXE analysis of Zn enzymes. Nuclear Instruments and Methods in Physics Research B, 150, 222-225. https://doi.org/10.1016/S0168-583X(98)01012-X
  • Söyüt, H., Beydemir, S., & Hisar, O. (2008). Effects of some metals on carbonic anhydrase from brains of rainbow trout. Biological Trace Element Research, 123, 179-190. https://doi.org/10.1007/s12011-008-8108-9
  • Supuran, C.T., & Scozzafava, A. (2001). Carbonic anhydrase ınhibitors. Current Medicinal Chemistry, 1, 61-97. https://doi.org/10.1002/med.10025
  • Supuran, C.T., Scozzafava, A., & Casini, A. (2003) Carbonic anhydrase inhibitors. Medical Research Reviews, 23, 146-157.
  • Şentürk, M., Gülçin, İ., Daştan, A., Küfrevioğlu, Ö.İ., & Supuran, C.T. (2009). Carbonic anhydrase inhibitors. Inhibition of human erythrocyte isozymes I and II with a series of antioxidant phenols. Bioorganic and Medicinal Chemistry, 17, 3207-3211. https://doi.org/10.1016/j.bmc.2009.01.067
  • Tsuzuki, M., & Miyachi, S. (1989). The function of carbonic anhydrase in aquatic photosynthesis. Aquatic Botany, 34, 85-104. https://doi.org/10.1016/0304-3770(89)90051-X
  • Wistrand, P.J. (2002). Carbonic anhydrase III in liver and muscle of male rats purification and properties. Upsala Journal of Medical Sciences, 107(2), 77-88.

Kalkan balığı (Psetta Maxima) kas dokusundan karbonik anhidrazın saflaştırılması, karakterizasyonu ve kinetik özelliklerinin belirlenmesi

Yıl 2023, Cilt: 1 Sayı: 1, 1 - 7, 18.06.2023

Öz

Bu çalışmada öncelikle kalkan (Psetta maxima) kas dokusundan karbonik anhidrazın saflaştırılması, enzimin kinetik davranışının analizi ve bazı özellikleri belirlenmiştir.Saflaştırma basamakları; hemolizat hazırlanması ve Sepharose-4B-L tirozin-sülfanilamid afinite jel kromatografisi işlemidir. CA enziminin saflaştırılmasında, enziminin spesifik aktivitesi 755.2 EU/mg protein, verim 69.05 ve saflaştırma katsayısı 50.65 olarak belirlenmiştir ve yapılan saflaştırma işlemlerinin hepsi +4ºC'de gerçekleşmiştir. SDS poliakrilamid jel elektroforezi ile saflaştırılan CA enziminin molekül ağırlığı 29.7 kDa olarak belirlenmiştir. Kalkan balığı (Psetta maxima) kas dokusundan saflaştırılan CA enziminin optimum iyonik şiddeti 0.020 M Tris-SO4 tamponu, optimum pH’sı 8.0, stabil pH’sı 8.0 ve optimum sıcaklığı 30ºC olarak gözlenmiştir. Enzimin kinetik özellikleri içerisinde yer alan KM ve Vmax değerleri belirlenmiştir.

Kaynakça

  • Badger, M.R., & Price, G.D. (1994). The role of carbonic anhydrase in photosynthesis. Annual Review of Plant Physiology and Plant Molecular Biology, 45, 369-392. https://doi.org/10.1146/annurev.pp.45.060194.002101
  • Bayram, E., Senturk, M., Kufrevioglu, O.I., & Supuran, C.T. (2008). In vitro inhibition of salicylic acid derivatives on human cytosolic carbonic anhydrase isozymes I and II. Bioorganic & Medicinal Chemistry, 16(20), 9101-9105. https://doi.org/10.1016/j.bmc.2008.09.028
  • Bradford, M.M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248. https://doi.org/10.1016/0003-2697(76)90527-3
  • Ceyhun, S.B., Senturk, M., Yerlikaya, E., Erdogan, O., Kufrevioglu, O.I., & Ekinci, D. (2011). Purification and characterization of carbonic anhydrase from the teleost fish Dicentrarchus labrax (European seabass) liver and toxicological effects of metals on enzyme activity. Environmental Toxicology and Pharmacology, 32(1), 69-74. https://doi.org/10.1016/j.etap.2011.03.013
  • Chegwidden, W.R., Edwards, Y., & Carter, N. (2000). The Carbonic Anhydrase-New Horizons. Molecular Bases of Inherited Disease (Scriver, C.R., Beaudet, A. L., Sly, W. S., and Valle, D., eds) 8th Ed., pp. 2165-2204, McGraw-Hill, Inc., Newyork.
  • Clare, B.W., & Supuran, C.T. (2000). Carbonic Anhydrase Inhibitors. Part 86. A QSAR study on some sulfonamide drugs which lower intra-ocular pressure, using the ACE non-linear statistical method. The European Journal of Medicinal Chemistry, 35, 859-865. https://doi.org/10.1016/S0223-5234(00)00182-3
  • Del Prete, S., Vullo, D., De Luca, V., Carginale, V., Osman, S.M., AlOthman, Z., Supuran, C.T., & Capasso, C. (2016). Cloning, expression, purification and sulfonamide inhibition profile of the complete domain of the η-carbonic anhydrase from Plasmodium falciparum. Bioorganic & Medicinal Chemistry Letters, 26(17), 4184-90. https://doi.org/10.1016/j.bmcl.2016.07.060
  • Demirdağ, R., Çomaklı, V., Şentürk, M., Ekinci, D., İrfan Küfrevioğlu, Ö., & Supuran, C.T. (2013). Purification and characterization of carbonic anhydrase from sheep kidney and effects of sulfonamides on enzyme activity. Bioorganic & medicinal chemistry, 21(6), 1522-1525. https://doi.org/10.1016/j.bmc.2012.08.018
  • Ekinci, D., Cavdar, H., Talaz, O., Senturk, M., & Supuran, C.T. (2010). NO-releasing esters show carbonic anhydrase inhibitory action against human isoforms I and II. Bioorganic & Medicinal Chemistry, 18(10), 3559-3563. https://doi.org/10.1016/j.bmc.2010.03.082
  • FAO (2010). The State of World Fisheries and Aquaculture. http://www.fao.org/docrep/013/i1820e/i1820e.pdf Göçer, H., Akıncıoğlu, A., Göksu, S., Gülcin, I., & Supuran, C.T. (2015). Carbonic anhydrase and acetylcholine esterase inhibitory effects of carbamates andsulfamoylcarbamates. Journal of enzyme inhibition and medicinal chemistry, 30(2), 316-320. https://doi.org/10.3109/14756366.2014.928704
  • Göksu, S., Naderi, A., Akbaba, Y., Kalın, P., Akıncıoglu, A., Gulcin, I., Durdagi, S., & Salmas, R.E. (2014). Carbonic anhydrase inhibitory properties of novelbenzylsulfamides using molecular modeling and experimental studies. Bioorganic chemistry, 56, 75-82. https://doi.org/10.1016/j.bioorg.2014.07.009
  • Graham, D., Reed, M.L., Patterson, B.D., Hockley, D.G., & Dwyer, M. (1984). Chemical properties, distribution and physiology of plant and algal carbonic anhydrases. Annals of the New York Academy of Sciences, 429, 222-237. https://doi.org/10.1111/j.1749-6632.1984.tb12340.x
  • Hara, S. (2001). Karadeniz kalkan balıgından (Psetta maxima) döllenmis yumurta ve larva elde edilmesi. SÜMAE, Yunus Arastırma Bülteni, 1,1.
  • Henry, R.P., Smatresk, N.J., & Cameron, J.N. (1988). The distribition of brachial carbonic anhydrase and the effect of gill and erythrocyte carbonic anhydrase inhibition in the cannel catfish, Ictalurus punctatus. Journal of Experimental Biology, 134, 201-218. https://doi.org/10.1242/jeb.134.1.201
  • Jo, B.H., & Hwang, I.S. (2019). Characterization and High-Level Periplasmic Expression of Thermostable α-Carbonic Anhydrase from Thermosulfurimonas Dismutans in Escherichia Coli for CO2 Capture and Utilization. International Journal of Molecular Sciences, 21(1), 103. https://doi.org/10.3390/ijms21010103
  • Jones, A. (1974). Sexual Maturity, Fecundity and Growth of the Turbot (Scophthalmus maximus L.). Journal of the Marine Biological Association of the United Kingdom, 54, 109-125. https://doi.org/10.1017/S0025315400022104
  • Kaya, E.D., Söyüt, H., & Beydemir, S. (2013). Carbonic anhydrase activity from thegilthead sea bream (Sparus aurata) liver: the toxicological effects of heavymetals. Environmental Toxicology and Pharmacology, 36, 514-521. https://doi.org/10.1016/j.etap.2013.05.019
  • Kucuk, M., & Gulcin, İ. (2016). Purification and characterization of the carbonic anhydrase enzyme from Black Sea trout (Salmo trutta Labrax Coruhensis) kidney and inhibition effects of some metal ions on enzyme activity. Environmental Toxicology and Pharmacology, 44, 134-9. https://doi.org/10.1016/j.etap.2016.04.011
  • Laemmli, D.K. (1970). Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature, 227, 680.
  • Maheshwari, N., Kumar, M., Thakur, I.S., & Srivastava, S. (2019). Cloning, expression and characterization of β- and γ‑carbonic anhydrase from Bacillus sp. SS105 for biomimetic sequestration of CO2. International journal of biological macromolecules, 131, 445-452. https://doi.org/10.1016/j.ijbiomac.2019.03.082
  • Memis, D. (2010). Deniz Balıkları Yetistiriciligi. Filiz Kitabevi, İstanbul, 152-172.
  • Munroe, T.A. (2005). Systematic Diversity of the Pleuronectiformes in: Flatfishes
  • Öztürk Sarikaya, S.B., Topal, F., Şentürk, M., Gülçin, İ., & Supuran, C.T. (2011). In vitro inhibition of a-carbonic anhydrase isozymes by some phenolic compounds. Bioorganic & Medicinal Chemistry Letters, 21, 4259. https://doi.org/10.1016/j.bmcl.2011.05.071
  • Peterson, R.E., Tu, C., & Linser, P.J. (1997). Isolation and characterisation of a carbonicanhyrdase homologue from the zebrafish (Danio rerio). Journal of Molecular Evolution, 448, 432-439. https://doi.org/10.1007/pl00006163
  • Rizzello, A., Ciardiello, M.A., Acierno, R., Carratore, V., Verri, T., Di Prisco, G., Storelli, C., & Maffia, M. (2007). Biochemical characterization of a s-glutathionylatedcarbonic anhydrase isolated from gills of the antarctic icefish. The Protein Journal, 26, 335-348. https://doi.org/10.1007/s10930-007-9076-1
  • Samsun, N. (2004). Sinop yöresinde avlanan kalkan (Scophthalmus maeoticus Pallas,1811) balıklarının bazı biyolojik özelliklerinin belirlenmesi üzerine bir arastırma. Ondokuz Mayıs Üniversitesi Fen Bilimleri Enstitüsü Su Ürünleri Avlama ve İşleme Teknolojisi Anabilim Dalı, Doktora Tezi, 171s., Samsun.
  • Segel, I.H. (1968). Biochemical calculations, second edition, Publised by, John Wiley and Sons Inc. New York, USA, 458 pp., ISBN: 0-471-77421-9
  • Sevgili, H., & Nezaki, G. (2010). Turbot Culture in France and Spain. Special Publication 5. February Emre Y. (Eds).
  • Solis, C., Olivera, A., Andrade, E., Ruvalcaba-Sil, J.L., Romero, I., & Celis, H. (1999). PIXE analysis of Zn enzymes. Nuclear Instruments and Methods in Physics Research B, 150, 222-225. https://doi.org/10.1016/S0168-583X(98)01012-X
  • Söyüt, H., Beydemir, S., & Hisar, O. (2008). Effects of some metals on carbonic anhydrase from brains of rainbow trout. Biological Trace Element Research, 123, 179-190. https://doi.org/10.1007/s12011-008-8108-9
  • Supuran, C.T., & Scozzafava, A. (2001). Carbonic anhydrase ınhibitors. Current Medicinal Chemistry, 1, 61-97. https://doi.org/10.1002/med.10025
  • Supuran, C.T., Scozzafava, A., & Casini, A. (2003) Carbonic anhydrase inhibitors. Medical Research Reviews, 23, 146-157.
  • Şentürk, M., Gülçin, İ., Daştan, A., Küfrevioğlu, Ö.İ., & Supuran, C.T. (2009). Carbonic anhydrase inhibitors. Inhibition of human erythrocyte isozymes I and II with a series of antioxidant phenols. Bioorganic and Medicinal Chemistry, 17, 3207-3211. https://doi.org/10.1016/j.bmc.2009.01.067
  • Tsuzuki, M., & Miyachi, S. (1989). The function of carbonic anhydrase in aquatic photosynthesis. Aquatic Botany, 34, 85-104. https://doi.org/10.1016/0304-3770(89)90051-X
  • Wistrand, P.J. (2002). Carbonic anhydrase III in liver and muscle of male rats purification and properties. Upsala Journal of Medical Sciences, 107(2), 77-88.
Toplam 35 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Konular Yapısal Biyoloji , Balık Yetiştiriciliği
Bölüm Araştırma Makalesi
Yazarlar

Ahmet Topal 0000-0002-7484-2599

Tuğrul Kurbanoğlu Bu kişi benim 0000-0001-8137-4765

Erken Görünüm Tarihi 13 Haziran 2023
Yayımlanma Tarihi 18 Haziran 2023
Gönderilme Tarihi 22 Mayıs 2023
Yayımlandığı Sayı Yıl 2023 Cilt: 1 Sayı: 1

Kaynak Göster

APA Topal, A., & Kurbanoğlu, T. (2023). Purification, characterization and dertermination of kinetic features of carbonic anhydrase from turbot (Psetta Maxima) muscle tissue. The Trout Journal of Atatürk University, 1(1), 1-7.

The Trout Journal of Atatürk University (Atatürk Üniversitesi Alabalık Dergisi)

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