TY - JOUR T1 - Anoxybacillus thermarum A4 Suşundaki Katalaz Aktivitesinin İncelenmesi ve Tam Hücre İmmobilizasyonu TT - Investigation of Catalase Activity from Anoxybacillus thermarum A4 strain and Whole Cell Immobilization AU - Dinçer, Barbaros AU - Durmaz, Murat AU - Adıgüzel, Ahmet PY - 2019 DA - December Y2 - 2019 DO - 10.35229/jaes.647364 JF - Journal of Anatolian Environmental and Animal Sciences JO - JAES PB - Bülent VEREP WT - DergiPark SN - 2548-0006 SP - 581 EP - 588 VL - 4 IS - 3 LA - tr AB - Yapılan bu çalışma ile Anoxybacillus thermarum A4 suşunun, katalaz enzimini üretebilme kapasitesi ve üretilen enzimin bazı kinetik verileri incelendi. Bununla birlikteA. thermarumA4 suşunun tam hücre immobilizasyonu agar ve agaroz ortamlarında gerçekleştirilerek, katalaz aktivitesi karakterize edildi. A4 suşunun yüksek katalaz aktivitesine sahip olduğu belirlendi. A4 suşu katalazının en yüksek aktivitesini pH 7,0 ve 50oC’de gösterdiği belirlendi. A4 suşu özütü katalazının 17,5±2,2 mM Kmdeğerine ve 250 000±980 U mg protein-1Vmaksdeğerine sahip olduğu belirlenirken, tam hücre immobilizasyonu sonrası Kmdeğeri 200±28 mM ve Vmaksdeğeri 50.000±413 U g jel-1olarak belirlendi. A. thermarum A4 suşundan elde edilen katalazın Sodyum azid, Potasyum siyanür, Civa (II) klorür ve 3-amino-1,2,4-triazol gibi bilinen katalaz inhibitörleri ile inhibe olduğu belirlendi. Bu çalışma sonucunda, materyal olarak kullanılan A. thermarum A4 suşu yüksek bir katalaz aktivitesine sahip olmasından dolayı hidrojen peroksidin kullanıldığı endüstri alanları için bir potansiyel katalaz kaynak olabileceği öngörüldü. KW - Katalaz KW - Hidrojen Peroksit Oksidoredüktaz KW - Termofilik Bakteri N2 - In this study, the capacity of Anoxybacillusthermarum A4 strain to produce the catalase enzyme and some kinetic data of theproduced enzyme were investigated. In addition, A. thermarum A4 strain was performed immobilization of its wholecell in agar and agarose environments, and their catalase activities werecharacterized. It was determined that A4 strain had high catalase activity. Thehighest activity of A4 strain catalase was determined at pH 7.0 and 50 oC.While catalase of A4 strain extract was determined to have 17.5 ± 2.2 mM Km and250,000 ± 980 U mg protein-1 Vmax,after whole cell immobilization , Km value was determined as 200 ± 28 mMand Vmax was 50,000 ± 413 U g gel-1. The catalase obtained from A.thermarum A4 strain was inhibited by known catalase inhibitors such as Sodiumazide, Potassium cyanide, Mercury (II) Lys and 3-amino-1,2,4-triazole. As aresult of this study, it was envisaged that the A. thermarum A4 strain used as material could be a potentialcatalase source for the industrial areas where hydrogen peroxide was usedbecause of its high catalase activity. CR - Aebi, H., 1984. Catalase in vitro, Methods in Enzymology, 105, 121-126. CR - Akgöl, S., Kaçar, Y., Özkara, S., Yavuz, H., Denizli, A. and Arica, MY., 2001. İmmobilization of Catalase Via Adsorption onto L-histidine Grafted Functional pHEMA Based Membran, Journal of Molecular Catalysis B: Enzymatic, 15, 197-206. CR - Brown-Peterson, NJ., Salin, ML., 1993. Purification of Catalase-Peroxidase from Halobacterium halobium: Characterization of Some Unique Properties of the Halophilic Enzyme, Journal of Bacteriology, 175 (13), 4197-4202. CR - Brown-Peterson, NJ., Salin, ML., 1995. Purification and Characterization of Mesohalic Catalase from Halophilic Bacterium Halobacterium halobium, Journal of Bacteriology, 177 (2), 378-384. CR - Costa, SA., Tzanov, T., Paar, A., Gudelj, M., Gübitz, GM., Cavaco-Paulo, A., 2001. Immobilization of Catalases from Bacillus SF on Alumina for the Treatment of Textile Bleaching Effluents, Enzyme and Microbial Technology, 28, 815-819. CR - Dinçer, B., 2005. Bazı Termofilik Bakterilerdeki Katalaz Aktivitesinin İncelenmesi, Karadeniz Teknik Üniversitesi Fen Bilimleri Enstitüsü, Doktora, Trabzon, Türkiye, 100s. CR - Durmaz, M., 2012. Anoxybacillus gonensis Z4 suşundaki katalaz aktivitesinin incelenmesi ve tam hücre immobilizasyonu, Recep Tayyip Erdoğan Üniversitesi Fen Bilimleri Enstitüsü. Yüksek Lisans, Rize, Türkiye, 75s. CR - Gonçalves, VM., de Cerqueira Leite, LC., Raw, I., Cabrera-Crespo, J., 1999. Purification of Catalase from Human Placenta, Biotechnology and Applied Biochemistry, 29, 73-77. CR - Grigoras, AG., 2017. Catalase Immobilization-A Review, Biochemical Engineering Journal, 117, 1-20. CR - Hatchikian, EC., LeGall, J., Bruschi, M., Dubourdieu, M., 1972. Regulation of the Reduction of Sulfite and Thiosulfate by Ferredoxin, Flavodoxin and Cytochrome C3 in Extracts of the Sulfate Reducer Desulfovibrio gigas, Biochimica et Biophysica Acta, 258, 701–708. CR - Hidalgo, A., Betancor, L., Mateo, C., Lopez-Gallego, F., Moreno, R., Berenguer, J., Guisan, JM., Fernández-Lafuente, R., 2004. Purification of a Catalase from Thermus thermophilus via IMAC Chromatography: Effect of the Support, Biotechnology Progress, 20, 1578-1582. CR - Hildebrandt, AG., Roots, I., 1975. Reduced Nicotinamide Adenine Dinucleotide Phosphate (NADPH)-dependent Formation and Breakdown of Hydrogen Peroxide Daring Mixed Function Oxidation Reaction in Liver Microsomes, Archives of Biochemistry and Biophysics, 171, 385-397. CR - Hillenbrand, T., 1999. Die abwasser situation in der deutschen papier-, textile-, und lederindustrie, GWF, Wasser/Abwasser, 140 (4), 267-273. CR - Kourkoutas, Y., Bekatorou, A., Banatb, IM., Marchant, R., Koutinas, AA., 2004. Immobilization technologies and support materials suitable inalcohol beverages production: a review. Food Microbiology, 2, 377-397. CR - Lowry, OH., Rosebrough, NJ., Farr, AL., Randall, RJ., 1951. Protein Measurement with the Folin Phenol Reagent, The Journal of Biological Chemistry, 193, 265-275. CR - Monti, D., Baldaro, E., Riva, S., 2003. Separation and Characterization of Two Catalase Activities İsolated from the Yeast Trigonopsis variabilis, Enzyme and Microbial Technology, 32, 596-605. CR - Terzenbach, DP., Blaut, M., 1998. Purification and Characterization of a Catalase from the Nonsulfur Phototrophic Bacterium Rhodobacter sphaeroides ATH 2.4.1 and Its Role in the Oxidative Stres Response, Archives of Microbiology, 169, 503-508. CR - Tischer, W., Wedekind, F., 1999. Immobilized Enzymes: Methods and Applications. Biocatalysis-From Discovery to Application, Springer, No: 200, s. 95-126, Berlin-Almanya. CR - Vatsyayan, P., Goswami, P., 2016. Highly Active and Stable Large Catalase Isolated from a Hydrocarbon Degrading Aspergillus terreus MTCC 6324. Enzyme Research, 1-8. CR - Wang, H., Tokusige, Y., Shinoyama, H., Fujii, T., Urakami, T., 1998. Purification and Characterization of a Thermostable Catalase from Culture Broth of Thermoascus aurantiacus. Journal of Fermentation and Bioengineering, 85 (2), 169-173. CR - Wang, H., Wang, J., Wang, J., Zhu, R., Shen, Y., Xu, Q., 2017. Spectroscopic method for the detection of 2,4-dichlorophenoxyacetic acid based on its inhibitory effect towards catalase immobilized on reusable magnetic Fe3O4-chitosan nanocomposite. Sensors and Actuators B: Chemical, 247, 146-154. CR - Wayne, LG., Diaz, GA., 1986. A Double Staining Method for Differentiating Between Two Classes of Mycobacterial Catalase in Polyacrylamide Electrophoresis Gels, Analytical Biochemistry, 157, 89–92. CR - Weck, M., 1991. Hydrogen peroxide-an environmentally acceptable textile bleaching agent, Text Praxis International, 2, 144-147. CR - Woodbury, W., Spencer, AK., Stahmann, MA., 1971. An Improved Procedure Using Ferricyanide for Detecting Catalase Isozymes. Analytical Biochemistry, 44, 301-305. CR - Zou, P., Schrempf, H., 2000. The Heme-Independent Manganese-Peroxidase Activity Depends on the Presence of the C-Terminal Domain within the Streptomyces reticuli Catalase-Peroxidase CpeB. European Journal of Biochemistry, 267, 2840-2849. UR - https://doi.org/10.35229/jaes.647364 L1 - https://dergipark.org.tr/en/download/article-file/906955 ER -