Purification and characterization Catalase from Bacillus subtilis PY79
Main Article Content
Abstract
Catalase is an enzyme present in peroxisome in most aerobic cells and is a central component of detoxifying processes that rapidly suppress the formation of H2O2 by catalysing the degradation of H2O2 into H2O and O2. In this study, catalase was purified from Bacillus subtilis PY79 by three steps: ammonium sunfate precipitation, anion exchange chromatography on Q-sepharose and cation exchange chromatography on CM-sepharose. The purified catalase showed high specific activity of 30717,2 U/mg with 8,89% yield. The enzyme remained active over a broad pH range of 5 – 11, with a peak in activity at pH 7,0. A broad optimum temperature range from 4 – 40oC was observed with highest activity at 37oC. The apparent Km for H2O2 was 14,4 mM and its Vmax was 16294,83 U/mg. Moreover, it was inhibited by NaN3, FeCl3, FeSO4, NaCl at concentrations of 5 μM, 15 mM, 50 mM, 2 M, respectively and MgSO4 or MnSO4 at 1 M was no inhibitory effect on this enzyme
Keywords:
Bacillus subtilis PY79, catalase, characterization, purification
References
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[19] Fusho Y., Yajima Y., Catalase from Bacillus subtilis IAM 1026 (Ferm BP-4844), United Sates Patent 5486467 (1996).
[20] Hussein AA., Purification and characterziation of thermo-alkali stable catalase from Bacillus sp, International Research Journal Biotechnology 3 (2012) 207.
[21] Suryani, Ambarsari L., Lindawati E., Isolation , Fractionation and Characterization of Catalase from Neurospora crassa (InaCC F226), Conference Series: Earth and Environmental Science 58 (2017) 012068.
[22] Loewen PC., Switala J., Purification and characterization of catalase-1 from Bacillus subtilis, Biochemistry and Cell Biology 65 (1987) 939
[23] Santoso P, Ambarsari L, Suryani, Yopi, Purification and Characterization of Catalase from Indigenous Fungi of Neurospora crassa InaCC F226, International Journal on Advanced Science Engineering Information Technology 6 (2016) 2088.
[2] Shin DH., Choi YS., Cho YH., Unusual properties of catalase a (Kat) of Pseudomonas aeruginosa Pa14 are associatedwith its biofilm peroxide resistance, Journal of Bacteriology 190 (2008) 2663.
[3] Aydemir T., Kuru K., Purification and partial characterization of catalase from chicken erythrocytes and the effect of various inhibitors an enzyme activity, Turkish Journal of Chemistry 27 (2003) 85.
[4] Nadeem MS., Khan IA., Murtaza BN., Muhammad K., Rauf A., Purrification and properties of liver catalase from water buffalo (Budalus bubalis), South Asian Journal of Life Sciences 32 (2015) 51.
[5] Vuillaume M., Reduced oxigen species, mutation induction and cancer initiation, Mutation Research 186 (1987) 43.
[6] Miyamoto T., Hayashi M., Takeuchi A., Okamoto T., Kawashima S., Takii T., Ayashi H., Onozaki K., Identification of a novel growwth-promoting factor with a wide target cell specturm from various tumor cells as catalase, Journal of Boichemistry (Tokyo) 120 (1996) 725.
[7] Esch F., Lin KI., Hills A., Zaman K., Baraban JM., Purification of a multipotent antideath activity from bovine liver and its identification as arginase: nitric oxide-independent inhibition of neuronal apoptosis, Journal of Neuroscience 18 (1998) 4083.
[8] Yabuki M., Kariya S., Ishisaka R., Yasuda T., Yoshioka T., Horton AA., Utsumi K., Resistance to nitric oxide-mediated apoptosis in HL-60 variant cells is associated with increased activities of Cu, Zn-superoxide dismutase and catalase, Free Radical Biology and Medicine 26 (1999) 325.
[9] Halliwell B., Gutteridge JM., Oxigen toxicity, oxigen radicals, transition metals and disease, Biochemical Journal 219 (2011) 1.
[10] Siwale RC., Yeboah GK., Addo R., Oettinger CW., D’Souza MJ., The effect of intracellilar antioxidant delivery (catalase) on hydrogen peroxide and pro-inflammatory cytokine synthesis: A new therapeutic horizon, Journal of Drug Target 17 (2009) 710.
[11] Robinet P., Wang Z., Hazen SL., Smith JD., A simple and sensitive enzymatic method for cholesterol quantification in macrophages and foam cells, Journal of Lipid Research 51 (2010) 3364.
[12] Wood JM., Decker H., Hartmann H., Senile hair graying: H2O2-mediated oxidative stress affects human hair color by blunting methionine sulfoxide repair, FASEB Journal 23 (2009) 2065.
[13] Yumoto I., Ichihasi D., Iwata H., Istokivics A., Ichise N., Matuyama H., Okuyama H., Kosei K., Purification and Characterization of Catalase from the Facultatively Psychrophilic Bacterium Vibriorumoiensis S-1T Exhibiting High Catalase Activity, Journal of Bacteriology 182 (2000) 1903.
[14] Zámocký M., Godočíková J., Gašperík J., Koller F., Polek B., Expression, purification, and sequence analysis of catalase-1 from the soil bacterium Comamonas terrigena NH3, Protein expression and purification 36 (2004) 115.
[15] Hossain SM., Nakayama M., Uchiyama H., Nakajima-Kambe T., Purification and Molecular Cloning of Catalase from Rhizobium radiobacter Strain 2-1, Journal of Environmental Biotechnology 8 (2008) 35.
[16] Hong HA., Huang JM., Khaneja R., Hiep LV., Urdaci MC., Cutting SM., The safety of Bacillus subtilis and Bacillus indicus as food probiotics, Journal of Applied Microbiology 105 (2008) 510.
[17] Lefevre M., Racedo SM., Denayrolles M., Ripert G., Desfougères T., Lobach AR., Simon R., Pélerin F., Jüsten P., Urdaci MC., Safety assessment of Bacillus subtilis CU1 for use a probiotic in husman, Regulatory Toxicology and Pharmacology 83 (2017) 54.
[18] Võ Thị Mai Hương, Trần Thanh Phong, Một số đặc trưng hóa sinh và khả năng kháng khuẩn của dịch chiết quả nhàu (Morinda citrifolia L.), Hội nghị khoa học toàn quốc về sinh thái và tài nguyên sinh vật lần thứ 5 (2006) 1073.
[19] Fusho Y., Yajima Y., Catalase from Bacillus subtilis IAM 1026 (Ferm BP-4844), United Sates Patent 5486467 (1996).
[20] Hussein AA., Purification and characterziation of thermo-alkali stable catalase from Bacillus sp, International Research Journal Biotechnology 3 (2012) 207.
[21] Suryani, Ambarsari L., Lindawati E., Isolation , Fractionation and Characterization of Catalase from Neurospora crassa (InaCC F226), Conference Series: Earth and Environmental Science 58 (2017) 012068.
[22] Loewen PC., Switala J., Purification and characterization of catalase-1 from Bacillus subtilis, Biochemistry and Cell Biology 65 (1987) 939
[23] Santoso P, Ambarsari L, Suryani, Yopi, Purification and Characterization of Catalase from Indigenous Fungi of Neurospora crassa InaCC F226, International Journal on Advanced Science Engineering Information Technology 6 (2016) 2088.