Anoxybacillus thermarum A4 Suşundaki Katalaz Aktivitesinin İncelenmesi ve Tam Hücre İmmobilizasyonu

Yıl 2019, Cilt: 4 Sayı: 3, 581 - 588, 30.12.2019

Barbaros Dinçer , Murat Durmaz Ahmet Adıgüzel

https://doi.org/10.35229/jaes.647364

Öz

Yapılan bu çalışma ile Anoxybacillus thermarum A4 suşunun, katalaz enzimini üretebilme kapasitesi ve üretilen enzimin bazı kinetik verileri incelendi. Bununla birlikte A. thermarum A4 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 50 ^oC’de gösterdiği belirlendi. A4 suşu özütü katalazının 17,5±2,2 mM K_m değerine ve 250 000±980 U mg protein^-1  V_maks değerine sahip olduğu belirlenirken, tam hücre immobilizasyonu sonrası K_m değeri 200±28 mM ve V_maks değeri  50.000±413 U g jel^-1 olarak 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.  

Anahtar Kelimeler

Katalaz, Hidrojen Peroksit Oksidoredüktaz, Termofilik Bakteri

Destekleyen Kurum

Recep Tayyip Erdoğan Üniversitesi

Investigation of Catalase Activity from Anoxybacillus thermarum A4 strain and Whole Cell Immobilization

Öz

In this study, the capacity of Anoxybacillus
thermarum A4 strain to produce the catalase enzyme and some kinetic data of the
produced enzyme were investigated. In addition, A. thermarum A4 strain was performed immobilization of its whole
cell in agar and agarose environments, and their catalase activities were
characterized. It was determined that A4 strain had high catalase activity. The
highest 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 and
250,000 ± 980 U mg protein^-1 Vmax, 
after whole cell immobilization , Km value was determined as 200 ± 28 mM
and 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 Sodium
azide, Potassium cyanide, Mercury (II) Lys and 3-amino-1,2,4-triazole. As a
result of this study, it was envisaged that the A. thermarum A4 strain used as material could be a potential
catalase source for the industrial areas where hydrogen peroxide was used
because of its high catalase activity.

Anahtar Kelimeler

Catalase, Hydrogen Peroxide Oxidoreductase, Thermophilic Bacteria

Kaynakça

• Aebi, H., 1984. Catalase in vitro, Methods in Enzymology, 105, 121-126.
• 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.
• 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.
• Brown-Peterson, NJ., Salin, ML., 1995. Purification and Characterization of Mesohalic Catalase from Halophilic Bacterium Halobacterium halobium, Journal of Bacteriology, 177 (2), 378-384.
• 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.
• Dinçer, B., 2005. Bazı Termofilik Bakterilerdeki Katalaz Aktivitesinin İncelenmesi, Karadeniz Teknik Üniversitesi Fen Bilimleri Enstitüsü, Doktora, Trabzon, Türkiye, 100s.
• 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.
• 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.
• Grigoras, AG., 2017. Catalase Immobilization-A Review, Biochemical Engineering Journal, 117, 1-20.
• 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.
• 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.
• 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.
• Hillenbrand, T., 1999. Die abwasser situation in der deutschen papier-, textile-, und lederindustrie, GWF, Wasser/Abwasser, 140 (4), 267-273.
• 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.
• Lowry, OH., Rosebrough, NJ., Farr, AL., Randall, RJ., 1951. Protein Measurement with the Folin Phenol Reagent, The Journal of Biological Chemistry, 193, 265-275.
• 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.
• 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.
• Tischer, W., Wedekind, F., 1999. Immobilized Enzymes: Methods and Applications. Biocatalysis-From Discovery to Application, Springer, No: 200, s. 95-126, Berlin-Almanya.
• Vatsyayan, P., Goswami, P., 2016. Highly Active and Stable Large Catalase Isolated from a Hydrocarbon Degrading Aspergillus terreus MTCC 6324. Enzyme Research, 1-8.
• 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.
• 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.
• 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.
• Weck, M., 1991. Hydrogen peroxide-an environmentally acceptable textile bleaching agent, Text Praxis International, 2, 144-147.
• Woodbury, W., Spencer, AK., Stahmann, MA., 1971. An Improved Procedure Using Ferricyanide for Detecting Catalase Isozymes. Analytical Biochemistry, 44, 301-305.
• 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.